516 lines
No EOL
80 KiB
XML
516 lines
No EOL
80 KiB
XML
<?xml version="1.0" encoding="utf-8"?>
|
||
<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
|
||
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">
|
||
|
||
<head><meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
|
||
<!-- AppResources meta begin -->
|
||
<meta name="paf-app-resources" content="" />
|
||
<script type="text/javascript">var ncbi_startTime = new Date();</script>
|
||
|
||
<!-- AppResources meta end -->
|
||
|
||
<!-- TemplateResources meta begin -->
|
||
<meta name="paf_template" content="" />
|
||
|
||
<!-- TemplateResources meta end -->
|
||
|
||
<!-- Logger begin -->
|
||
<meta name="ncbi_db" content="books" /><meta name="ncbi_pdid" content="book-part" /><meta name="ncbi_acc" content="NBK604916" /><meta name="ncbi_domain" content="nciprotocols" /><meta name="ncbi_report" content="classic" /><meta name="ncbi_type" content="fulltext" /><meta name="ncbi_objectid" content="" /><meta name="ncbi_pcid" content="/NBK604916/?report=classic" /><meta name="ncbi_pagename" content="Measurement of Nanoparticle Effects on Cytotoxic Activity of NK Cells by Label-Free RT-CES System - National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols - NCBI Bookshelf" /><meta name="ncbi_bookparttype" content="chapter" /><meta name="ncbi_app" content="bookshelf" />
|
||
<!-- Logger end -->
|
||
|
||
<title>Measurement of Nanoparticle Effects on Cytotoxic Activity of NK Cells by Label-Free RT-CES System - National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols - NCBI Bookshelf</title>
|
||
|
||
<!-- AppResources external_resources begin -->
|
||
<link rel="stylesheet" href="/core/jig/1.15.2/css/jig.min.css" /><script type="text/javascript" src="/core/jig/1.15.2/js/jig.min.js"></script>
|
||
|
||
<!-- AppResources external_resources end -->
|
||
|
||
<!-- Page meta begin -->
|
||
<meta name="robots" content="INDEX,FOLLOW,NOARCHIVE" /><meta name="citation_inbook_title" content="National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols [Internet]" /><meta name="citation_title" content="Measurement of Nanoparticle Effects on Cytotoxic Activity of NK Cells by Label-Free RT-CES System" /><meta name="citation_publisher" content="National Cancer Institute (US)" /><meta name="citation_date" content="2020/06" /><meta name="citation_author" content="Timothy M. Potter" /><meta name="citation_author" content="Edward Cedrone" /><meta name="citation_author" content="Barry W. Neun" /><meta name="citation_author" content="Marina A. Dobrovolskaia" /><meta name="citation_pmid" content="39013070" /><meta name="citation_doi" content="10.17917/GMXG-BH29" /><meta name="citation_fulltext_html_url" content="https://www.ncbi.nlm.nih.gov/books/NBK604916/" /><link rel="schema.DC" href="http://purl.org/DC/elements/1.0/" /><meta name="DC.Title" content="Measurement of Nanoparticle Effects on Cytotoxic Activity of NK Cells by Label-Free RT-CES System" /><meta name="DC.Type" content="Text" /><meta name="DC.Publisher" content="National Cancer Institute (US)" /><meta name="DC.Contributor" content="Timothy M. Potter" /><meta name="DC.Contributor" content="Edward Cedrone" /><meta name="DC.Contributor" content="Barry W. Neun" /><meta name="DC.Contributor" content="Marina A. Dobrovolskaia" /><meta name="DC.Date" content="2020/06" /><meta name="DC.Identifier" content="https://www.ncbi.nlm.nih.gov/books/NBK604916/" /><meta name="description" content="Natural killer (NK) cells are a type of lymphocyte which play a major role in the host-rejection of both cancer cells and cells infected by virus. NK cells carry small granules in their cytoplasm which contain special proteins such as perforin and granzymes. When NK cells release perforin in close proximity to target cells (i.e., tumorous or virus-infected cells), it forms pores in the cell membrane of the target cell through which the granzymes and associated molecules can enter, inducing apoptosis. Cytotoxic activity of NK cells is an important component of innate immunity which provides quick body response to cancerous or virus infected cells before more specialized adaptive immunity is generated. Understanding a drug’s effect on the cytotoxicity of NK cells is thus an important part of immunotoxicity studies aimed at identifying potential immunosuppression." /><meta name="og:title" content="Measurement of Nanoparticle Effects on Cytotoxic Activity of NK Cells by Label-Free RT-CES System" /><meta name="og:type" content="book" /><meta name="og:description" content="Natural killer (NK) cells are a type of lymphocyte which play a major role in the host-rejection of both cancer cells and cells infected by virus. NK cells carry small granules in their cytoplasm which contain special proteins such as perforin and granzymes. When NK cells release perforin in close proximity to target cells (i.e., tumorous or virus-infected cells), it forms pores in the cell membrane of the target cell through which the granzymes and associated molecules can enter, inducing apoptosis. Cytotoxic activity of NK cells is an important component of innate immunity which provides quick body response to cancerous or virus infected cells before more specialized adaptive immunity is generated. Understanding a drug’s effect on the cytotoxicity of NK cells is thus an important part of immunotoxicity studies aimed at identifying potential immunosuppression." /><meta name="og:url" content="https://www.ncbi.nlm.nih.gov/books/NBK604916/" /><meta name="og:site_name" content="NCBI Bookshelf" /><meta name="og:image" content="https://www.ncbi.nlm.nih.gov/corehtml/pmc/pmcgifs/bookshelf/thumbs/th-nciprotocols-lrg.png" /><meta name="twitter:card" content="summary" /><meta name="twitter:site" content="@ncbibooks" /><meta name="bk-non-canon-loc" content="/books/n/nciprotocols/ncipr54/" /><link rel="canonical" href="https://www.ncbi.nlm.nih.gov/books/NBK604916/" /><link rel="stylesheet" href="/corehtml/pmc/css/figpopup.css" type="text/css" media="screen" /><link rel="stylesheet" href="/corehtml/pmc/css/bookshelf/2.26/css/books.min.css" type="text/css" /><link rel="stylesheet" href="/corehtml/pmc/css/bookshelf/2.26/css/books_print.min.css" type="text/css" media="print" /><style type="text/css">p a.figpopup{display:inline !important} .bk_tt {font-family: monospace} .first-line-outdent .bk_ref {display: inline} .body-content h2, .body-content .h2 {border-bottom: 1px solid #97B0C8} .body-content h2.inline {border-bottom: none} a.page-toc-label , .jig-ncbismoothscroll a {text-decoration:none;border:0 !important} .temp-labeled-list .graphic {display:inline-block !important} .temp-labeled-list img{width:100%}</style><script type="text/javascript" src="/corehtml/pmc/js/jquery.hoverIntent.min.js"> </script><script type="text/javascript" src="/corehtml/pmc/js/common.min.js?_=3.18"> </script><script type="text/javascript" src="/corehtml/pmc/js/large-obj-scrollbars.min.js"> </script><script type="text/javascript">window.name="mainwindow";</script><script type="text/javascript" src="/core/mathjax/2.7.9/MathJax.js?config=/corehtml/pmc/js/mathjax-config-classic.3.4.js"></script><script type="text/javascript" src="/corehtml/pmc/js/bookshelf/2.26/book-toc.min.js"> </script><script type="text/javascript" src="/corehtml/pmc/js/bookshelf/2.26/books.min.js"> </script><meta name="book-collection" content="NONE" />
|
||
|
||
<!-- Page meta end -->
|
||
<link rel="shortcut icon" href="//www.ncbi.nlm.nih.gov/favicon.ico" /><meta name="ncbi_phid" content="CE8E2C587D840A210000000000F100D4.m_13" />
|
||
<meta name='referrer' content='origin-when-cross-origin'/><link type="text/css" rel="stylesheet" href="//static.pubmed.gov/portal/portal3rc.fcgi/4216699/css/3852956/3985586/3808861/4121862/3974050/3917732/251717/4216701/14534/45193/4113719/3849091/3984811/3751656/4033350/3840896/3577051/3852958/4008682/4207974/4206132/4062871/12930/3964959/3854974/36029/4128070/9685/3549676/3609192/3609193/3609213/3395586.css" /><link type="text/css" rel="stylesheet" href="//static.pubmed.gov/portal/portal3rc.fcgi/4216699/css/3411343/3882866.css" media="print" /></head>
|
||
<body class="book-part">
|
||
<div class="grid">
|
||
<div class="col twelve_col nomargin shadow">
|
||
<!-- System messages like service outage or JS required; this is handled by the TemplateResources portlet -->
|
||
<div class="sysmessages">
|
||
<noscript>
|
||
<p class="nojs">
|
||
<strong>Warning:</strong>
|
||
The NCBI web site requires JavaScript to function.
|
||
<a href="/guide/browsers/#enablejs" title="Learn how to enable JavaScript" target="_blank">more...</a>
|
||
</p>
|
||
</noscript>
|
||
</div>
|
||
<!--/.sysmessage-->
|
||
<div class="wrap">
|
||
<div class="page">
|
||
<div class="top">
|
||
<div id="universal_header">
|
||
<section class="usa-banner">
|
||
<div class="usa-accordion">
|
||
<header class="usa-banner-header">
|
||
<div class="usa-grid usa-banner-inner">
|
||
<img src="https://www.ncbi.nlm.nih.gov/coreutils/uswds/img/favicons/favicon-57.png" alt="U.S. flag" />
|
||
<p>An official website of the United States government</p>
|
||
<button class="non-usa-accordion-button usa-banner-button" aria-expanded="false" aria-controls="gov-banner-top" type="button">
|
||
<span class="usa-banner-button-text">Here's how you know</span>
|
||
</button>
|
||
</div>
|
||
</header>
|
||
<div class="usa-banner-content usa-grid usa-accordion-content" id="gov-banner-top" aria-hidden="true">
|
||
<div class="usa-banner-guidance-gov usa-width-one-half">
|
||
<img class="usa-banner-icon usa-media_block-img" src="https://www.ncbi.nlm.nih.gov/coreutils/uswds/img/icon-dot-gov.svg" alt="Dot gov" />
|
||
<div class="usa-media_block-body">
|
||
<p>
|
||
<strong>The .gov means it's official.</strong>
|
||
<br />
|
||
Federal government websites often end in .gov or .mil. Before
|
||
sharing sensitive information, make sure you're on a federal
|
||
government site.
|
||
</p>
|
||
</div>
|
||
</div>
|
||
<div class="usa-banner-guidance-ssl usa-width-one-half">
|
||
<img class="usa-banner-icon usa-media_block-img" src="https://www.ncbi.nlm.nih.gov/coreutils/uswds/img/icon-https.svg" alt="Https" />
|
||
<div class="usa-media_block-body">
|
||
<p>
|
||
<strong>The site is secure.</strong>
|
||
<br />
|
||
The <strong>https://</strong> ensures that you are connecting to the
|
||
official website and that any information you provide is encrypted
|
||
and transmitted securely.
|
||
</p>
|
||
</div>
|
||
</div>
|
||
</div>
|
||
</div>
|
||
</section>
|
||
<div class="usa-overlay"></div>
|
||
<header class="ncbi-header" role="banner" data-section="Header">
|
||
|
||
<div class="usa-grid">
|
||
<div class="usa-width-one-whole">
|
||
|
||
<div class="ncbi-header__logo">
|
||
<a href="/" class="logo" aria-label="NCBI Logo" data-ga-action="click_image" data-ga-label="NIH NLM Logo">
|
||
<img src="https://www.ncbi.nlm.nih.gov/coreutils/nwds/img/logos/AgencyLogo.svg" alt="NIH NLM Logo" />
|
||
</a>
|
||
</div>
|
||
|
||
<div class="ncbi-header__account">
|
||
<a id="account_login" href="https://account.ncbi.nlm.nih.gov" class="usa-button header-button" style="display:none" data-ga-action="open_menu" data-ga-label="account_menu">Log in</a>
|
||
<button id="account_info" class="header-button" style="display:none" aria-controls="account_popup" type="button">
|
||
<span class="fa fa-user" aria-hidden="true">
|
||
<svg xmlns="http://www.w3.org/2000/svg" viewBox="0 0 24 24" width="20px" height="20px">
|
||
<g style="fill: #fff">
|
||
<ellipse cx="12" cy="8" rx="5" ry="6"></ellipse>
|
||
<path d="M21.8,19.1c-0.9-1.8-2.6-3.3-4.8-4.2c-0.6-0.2-1.3-0.2-1.8,0.1c-1,0.6-2,0.9-3.2,0.9s-2.2-0.3-3.2-0.9 C8.3,14.8,7.6,14.7,7,15c-2.2,0.9-3.9,2.4-4.8,4.2C1.5,20.5,2.6,22,4.1,22h15.8C21.4,22,22.5,20.5,21.8,19.1z"></path>
|
||
</g>
|
||
</svg>
|
||
</span>
|
||
<span class="username desktop-only" aria-hidden="true" id="uname_short"></span>
|
||
<span class="sr-only">Show account info</span>
|
||
</button>
|
||
</div>
|
||
|
||
<div class="ncbi-popup-anchor">
|
||
<div class="ncbi-popup account-popup" id="account_popup" aria-hidden="true">
|
||
<div class="ncbi-popup-head">
|
||
<button class="ncbi-close-button" data-ga-action="close_menu" data-ga-label="account_menu" type="button">
|
||
<span class="fa fa-times">
|
||
<svg xmlns="http://www.w3.org/2000/svg" viewBox="0 0 48 48" width="24px" height="24px">
|
||
<path d="M38 12.83l-2.83-2.83-11.17 11.17-11.17-11.17-2.83 2.83 11.17 11.17-11.17 11.17 2.83 2.83 11.17-11.17 11.17 11.17 2.83-2.83-11.17-11.17z"></path>
|
||
</svg>
|
||
</span>
|
||
<span class="usa-sr-only">Close</span></button>
|
||
<h4>Account</h4>
|
||
</div>
|
||
<div class="account-user-info">
|
||
Logged in as:<br />
|
||
<b><span class="username" id="uname_long">username</span></b>
|
||
</div>
|
||
<div class="account-links">
|
||
<ul class="usa-unstyled-list">
|
||
<li><a id="account_myncbi" href="/myncbi/" class="set-base-url" data-ga-action="click_menu_item" data-ga-label="account_myncbi">Dashboard</a></li>
|
||
<li><a id="account_pubs" href="/myncbi/collections/bibliography/" class="set-base-url" data-ga-action="click_menu_item" data-ga-label="account_pubs">Publications</a></li>
|
||
<li><a id="account_settings" href="/account/settings/" class="set-base-url" data-ga-action="click_menu_item" data-ga-label="account_settings">Account settings</a></li>
|
||
<li><a id="account_logout" href="/account/signout/" class="set-base-url" data-ga-action="click_menu_item" data-ga-label="account_logout">Log out</a></li>
|
||
</ul>
|
||
</div>
|
||
</div>
|
||
</div>
|
||
|
||
</div>
|
||
</div>
|
||
</header>
|
||
<div role="navigation" aria-label="access keys">
|
||
<a id="nws_header_accesskey_0" href="https://www.ncbi.nlm.nih.gov/guide/browsers/#ncbi_accesskeys" class="usa-sr-only" accesskey="0" tabindex="-1">Access keys</a>
|
||
<a id="nws_header_accesskey_1" href="https://www.ncbi.nlm.nih.gov" class="usa-sr-only" accesskey="1" tabindex="-1">NCBI Homepage</a>
|
||
<a id="nws_header_accesskey_2" href="/myncbi/" class="set-base-url usa-sr-only" accesskey="2" tabindex="-1">MyNCBI Homepage</a>
|
||
<a id="nws_header_accesskey_3" href="#maincontent" class="usa-sr-only" accesskey="3" tabindex="-1">Main Content</a>
|
||
<a id="nws_header_accesskey_4" href="#" class="usa-sr-only" accesskey="4" tabindex="-1">Main Navigation</a>
|
||
</div>
|
||
<section data-section="Alerts">
|
||
<div class="ncbi-alerts-placeholder"></div>
|
||
</section>
|
||
</div>
|
||
<div class="header">
|
||
<div class="res_logo"><h1 class="res_name"><a href="/books/" title="Bookshelf home">Bookshelf</a></h1><h2 class="res_tagline"></h2></div>
|
||
<div class="search"><form method="get" action="/books/"><div class="search_form"><label for="database" class="offscreen_noflow">Search database</label><select id="database"><optgroup label="Recent"><option value="books" selected="selected" data-ac_dict="bookshelf-search">Books</option><option value="pubmed">PubMed</option><option value="medgen">MedGen</option><option value="gene" class="last">Gene</option></optgroup><optgroup label="All"><option value="gquery">All Databases</option><option value="assembly">Assembly</option><option value="biocollections">Biocollections</option><option value="bioproject">BioProject</option><option value="biosample">BioSample</option><option value="books" data-ac_dict="bookshelf-search">Books</option><option value="clinvar">ClinVar</option><option value="cdd">Conserved Domains</option><option value="gap">dbGaP</option><option value="dbvar">dbVar</option><option value="gene">Gene</option><option value="genome">Genome</option><option value="gds">GEO DataSets</option><option value="geoprofiles">GEO Profiles</option><option value="gtr">GTR</option><option value="ipg">Identical Protein Groups</option><option value="medgen">MedGen</option><option value="mesh">MeSH</option><option value="nlmcatalog">NLM Catalog</option><option value="nuccore">Nucleotide</option><option value="omim">OMIM</option><option value="pmc">PMC</option><option value="protein">Protein</option><option value="proteinclusters">Protein Clusters</option><option value="protfam">Protein Family Models</option><option value="pcassay">PubChem BioAssay</option><option value="pccompound">PubChem Compound</option><option value="pcsubstance">PubChem Substance</option><option value="pubmed">PubMed</option><option value="snp">SNP</option><option value="sra">SRA</option><option value="structure">Structure</option><option value="taxonomy">Taxonomy</option><option value="toolkit">ToolKit</option><option value="toolkitall">ToolKitAll</option><option value="toolkitbookgh">ToolKitBookgh</option></optgroup></select><div class="nowrap"><label for="term" class="offscreen_noflow" accesskey="/">Search term</label><div class="nowrap"><input type="text" name="term" id="term" title="Search Books. Use up and down arrows to choose an item from the autocomplete." value="" class="jig-ncbiclearbutton jig-ncbiautocomplete" data-jigconfig="dictionary:'bookshelf-search',disableUrl:'NcbiSearchBarAutoComplCtrl'" autocomplete="off" data-sbconfig="ds:'no',pjs:'no',afs:'no'" /></div><button id="search" type="submit" class="button_search nowrap" cmd="go">Search</button></div></div></form><ul class="searchlinks inline_list"><li>
|
||
<a href="/books/browse/">Browse Titles</a>
|
||
</li><li>
|
||
<a href="/books/advanced/">Advanced</a>
|
||
</li><li class="help">
|
||
<a href="/books/NBK3833/">Help</a>
|
||
</li><li class="disclaimer">
|
||
<a target="_blank" data-ga-category="literature_resources" data-ga-action="link_click" data-ga-label="disclaimer_link" href="https://www.ncbi.nlm.nih.gov/books/about/disclaimer/">Disclaimer</a>
|
||
</li></ul></div>
|
||
</div>
|
||
|
||
|
||
|
||
<!--<component id="Page" label="headcontent"/>-->
|
||
|
||
</div>
|
||
<div class="content">
|
||
<!-- site messages -->
|
||
<!-- Custom content 1 -->
|
||
<div class="col1">
|
||
|
||
</div>
|
||
|
||
<div class="container">
|
||
<div id="maincontent" class="content eight_col col">
|
||
<!-- Custom content in the left column above book nav -->
|
||
<div class="col2">
|
||
|
||
</div>
|
||
|
||
<!-- Book content -->
|
||
|
||
|
||
<!-- Custom content between navigation and content -->
|
||
<div class="col3">
|
||
|
||
</div>
|
||
|
||
<div class="document">
|
||
<div class="pre-content"><div><div class="bk_prnt"><p class="small">NCBI Bookshelf. A service of the National Library of Medicine, National Institutes of Health.</p><p>National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols [Internet]. Bethesda (MD): National Cancer Institute (US); 2005 May 1-. doi: 10.17917/GMXG-BH29</p></div><div class="iconblock clearfix whole_rhythm no_top_margin bk_noprnt"><a class="img_link icnblk_img" title="Table of Contents Page" href="/books/n/nciprotocols/"><img class="source-thumb" src="/corehtml/pmc/pmcgifs/bookshelf/thumbs/th-nciprotocols-lrg.png" alt="Cover of National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols" height="100px" width="80px" /></a><div class="icnblk_cntnt eight_col"><h2>National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols [Internet].</h2><a data-jig="ncbitoggler" href="#__NBK604916_dtls__">Show details</a><div style="display:none" class="ui-widget" id="__NBK604916_dtls__"><div>Bethesda (MD): <a href="https://www.cancer.gov/nano/research/ncl" ref="pagearea=page-banner&targetsite=external&targetcat=link&targettype=publisher">National Cancer Institute (US)</a>; 2005 May 1-.</div></div><div class="half_rhythm"><ul class="inline_list"><li style="margin-right:1em"><a class="bk_cntns" href="/books/n/nciprotocols/">Contents</a></li></ul></div><div class="bk_noprnt"><form method="get" action="/books/n/nciprotocols/" id="bk_srch"><div class="bk_search"><label for="bk_term" class="offscreen_noflow">Search term</label><input type="text" title="Search this book" id="bk_term" name="term" value="" data-jig="ncbiclearbutton" /> <input type="submit" class="jig-ncbibutton" value="Search this book" submit="false" style="padding: 0.1em 0.4em;" /></div></form></div></div><div class="icnblk_cntnt two_col"><div class="pagination bk_noprnt"><a class="active page_link prev" href="/books/n/nciprotocols/ncipr55/" title="Previous page in this title">< Prev</a><a class="active page_link next" href="/books/n/nciprotocols/ncipr49/" title="Next page in this title">Next ></a></div></div></div></div></div>
|
||
<div class="main-content lit-style" itemscope="itemscope" itemtype="http://schema.org/CreativeWork"><div class="meta-content fm-sec"><h1 id="_NBK604916_"><span class="label">NCL Method ITA-11</span><span class="title" itemprop="name">Measurement of Nanoparticle Effects on Cytotoxic Activity of NK Cells by Label-Free RT-CES System</span></h1><div class="subtitle whole_rhythm">Version 3</div><p class="contrib-group"><span itemprop="author">Timothy M. Potter</span>, <span itemprop="author">Edward Cedrone</span>, <span itemprop="author">Barry W. Neun</span>, and <span itemprop="author">Marina A. Dobrovolskaia</span>.</p><a data-jig="ncbitoggler" href="#__NBK604916_ai__" style="border:0;text-decoration:none">Author Information and Affiliations</a><div style="display:none" class="ui-widget" id="__NBK604916_ai__"><p class="contrib-group"><h4>Authors</h4><span itemprop="author">Timothy M. Potter</span>, <span itemprop="author">Edward Cedrone</span>, <span itemprop="author">Barry W. Neun</span>, and <span itemprop="author">Marina A. Dobrovolskaia</span><sup><img src="/corehtml/pmc/pmcgifs/corrauth.gif" alt="corresponding author" /></sup><sup>1</sup>.</p><h4>Contact</h4><div class="affiliation"><sup>1</sup> <span class="email-label">Email: </span><a href="mailto:dev@null" data-email="vog.hin.liam@aniram" class="oemail">vog.hin.liam@aniram</a></div><div class="affiliation"><sup>1</sup> Nanotechnology Characterization Lab, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research sponsored by the National Cancer Institute, Frederick, MD 21702</div><div><sup><img src="/corehtml/pmc/pmcgifs/corrauth.gif" alt="corresponding author" /></sup>Corresponding author.</div></div><p class="small">Published: <span itemprop="datePublished">June 2020</span>.</p></div><div class="jig-ncbiinpagenav body-content whole_rhythm" data-jigconfig="allHeadingLevels: ['h2'],smoothScroll: false" itemprop="text"><div id="ncipr54.s1"><h2 id="_ncipr54_s1_">1. Introduction</h2><p>Natural killer (NK) cells are a type of lymphocyte which play a major role in the host-rejection of both cancer cells and cells infected by virus. NK cells carry small granules in their cytoplasm which contain special proteins such as perforin and granzymes. When NK cells release perforin in close proximity to target cells (i.e., tumorous or virus-infected cells), it forms pores in the cell membrane of the target cell through which the granzymes and associated molecules can enter, inducing apoptosis. Cytotoxic activity of NK cells is an important component of innate immunity which provides quick body response to cancerous or virus infected cells before more specialized adaptive immunity is generated. Understanding a drug’s effect on the cytotoxicity of NK cells is thus an important part of immunotoxicity studies aimed at identifying potential immunosuppression.</p></div><div id="ncipr54.s2"><h2 id="_ncipr54_s2_">2. Principles</h2><p>This document describes a protocol for assessing the effect of nanoparticles on the capacity of human natural killer (NK) cells to lyse tumorous target cells under <i>in vitro</i> conditions. In this method, the NK92 cell line is used as the model for natural killer cells, and the hepatocellular carcinoma HepG2 cell line is used as the model for target cells. Viability of HepG2 cells following the addition of untreated or nanoparticle-treated NK92 cells is monitored in real time using a real-time cell electronic system (RT-CES) [<a class="bk_pop" href="#ncipr54.ref1">1</a>,<a class="bk_pop" href="#ncipr54.ref2">2</a>].</p></div><div id="ncipr54.s3"><h2 id="_ncipr54_s3_">3. Reagents, Materials, and Equipment</h2><blockquote><p><i>Note: The NCL does not endorse any of the suppliers listed below; these reagents were used in the development of the protocol and their inclusion is for informational purposes only. Equivalent supplies from alternate vendors can be substituted. Please note that suppliers may undergo a name change due to a variety of factors. Brands and part numbers typically remain consistent but may also change over time</i>.</p></blockquote><dl id="ncipr54.l1" class="temp-labeled-list"><dt>3.1.</dt><dd id="ncipr54.lt1"><p class="no_top_margin">Reagents
|
||
<dl id="ncipr54.l2" class="temp-labeled-list"><dt>3.1.1.</dt><dd id="ncipr54.lt2"><p class="no_top_margin">PBS (GE Life Science, Hyclone, SH30256.01)</p></dd><dt>3.1.2.</dt><dd id="ncipr54.lt3"><p class="no_top_margin">Fetal Bovine Serum (GE Life Sciences, Hyclone, SH30070.03)</p></dd><dt>3.1.3.</dt><dd id="ncipr54.lt4"><p class="no_top_margin">Horse Serum (GE Life Sciences, Hyclone, SH30074.03)</p></dd><dt>3.1.4.</dt><dd id="ncipr54.lt5"><p class="no_top_margin">MEM, Alpha Modification (GE Life Sciences, Hyclone, SH30568.01)</p></dd><dt>3.1.5.</dt><dd id="ncipr54.lt6"><p class="no_top_margin">RPMI-1640 (GE Life Sciences, Hyclone, SH30096.01)</p></dd><dt>3.1.6.</dt><dd id="ncipr54.lt7"><p class="no_top_margin">L-glutamine (GE Life Sciences, Hyclone, SH30034.01)</p></dd><dt>3.1.7.</dt><dd id="ncipr54.lt8"><p class="no_top_margin">Myo-inositol (Sigma-Aldrich, I7508)</p></dd><dt>3.1.8.</dt><dd id="ncipr54.lt9"><p class="no_top_margin">Folic Acid (Sigma-Aldrich, F8758)</p></dd><dt>3.1.9.</dt><dd id="ncipr54.lt10"><p class="no_top_margin">2-Mercaptoethanol (Gibco, 21985-023)</p></dd><dt>3.1.10.</dt><dd id="ncipr54.lt11"><p class="no_top_margin">Recombinant Human IL-2 (R&D Systems, 202-IL-010)</p></dd><dt>3.1.11.</dt><dd id="ncipr54.lt12"><p class="no_top_margin">Trypan Blue solution (Gibco, 15250-061)</p></dd></dl></p></dd><dt>3.2.</dt><dd id="ncipr54.lt13"><p class="no_top_margin">Materials
|
||
<dl id="ncipr54.l3" class="temp-labeled-list"><dt>3.2.1.</dt><dd id="ncipr54.lt14"><p class="no_top_margin">Pipettes ranging from 0.05 mL to 10 mL</p></dd><dt>3.2.2.</dt><dd id="ncipr54.lt15"><p class="no_top_margin">Flat bottom 16 well E-plates</p></dd><dt>3.2.3.</dt><dd id="ncipr54.lt16"><p class="no_top_margin">Polypropylene tubes, 5 and 15 mL</p></dd><dt>3.2.4.</dt><dd id="ncipr54.lt17"><p class="no_top_margin">Reagent reservoirs</p></dd><dt>3.2.5.</dt><dd id="ncipr54.lt18"><p class="no_top_margin">T25 culture flasks</p></dd></dl>
|
||
<blockquote><p><i>Note : Certain models of RT-CES instrument can operate with 96-well E-plates</i></p></blockquote></p></dd><dt>3.3.</dt><dd id="ncipr54.lt19"><p class="no_top_margin">Equipment
|
||
<dl id="ncipr54.l4" class="temp-labeled-list"><dt>3.3.1.</dt><dd id="ncipr54.lt20"><p class="no_top_margin">Centrifuge</p></dd><dt>3.3.2.</dt><dd id="ncipr54.lt21"><p class="no_top_margin">Refrigerator, 2-8ºC</p></dd><dt>3.3.3.</dt><dd id="ncipr54.lt22"><p class="no_top_margin">Freezer, −20ºC</p></dd><dt>3.3.4.</dt><dd id="ncipr54.lt23"><p class="no_top_margin">Cell culture incubator with 5% CO<sub>2</sub> and 95% humidity</p></dd><dt>3.3.5.</dt><dd id="ncipr54.lt24"><p class="no_top_margin">Biohazard safety cabinet approved for level II handling of biological material</p></dd><dt>3.3.6.</dt><dd id="ncipr54.lt25"><p class="no_top_margin">Inverted microscope</p></dd><dt>3.3.7.</dt><dd id="ncipr54.lt26"><p class="no_top_margin">Vortex</p></dd><dt>3.3.8.</dt><dd id="ncipr54.lt27"><p class="no_top_margin">Hemacytometer</p></dd><dt>3.3.9.</dt><dd id="ncipr54.lt28"><p class="no_top_margin">RT-CES instrument (ACEA Biosciences)</p></dd></dl></p></dd></dl></div><div id="ncipr54.s4"><h2 id="_ncipr54_s4_">4. Reagent Preparation</h2><dl id="ncipr54.l5" class="temp-labeled-list"><dt>4.1.</dt><dd id="ncipr54.lt29"><p class="no_top_margin">
|
||
<u>Heat-inactivation of fetal bovine serum</u>
|
||
</p><p>Thaw a bottle with FBS at room temperature or overnight at 2-8ºC and allow to equilibrate to room temperature. Incubate 30 m at 56ºC in a water bath, mixing every 5 minutes. Single use aliquots may be stored at 2-8ºC for up to one month or at a nominal temperature of −20ºC indefinitely.</p></dd><dt>4.2.</dt><dd id="ncipr54.lt30"><p class="no_top_margin">
|
||
<u>Complete RPMI medium (to maintain HepG2 cells)</u>
|
||
</p><p>The complete RPMI medium should contain the following reagents: 10% FBS (heat inactivated); 2 mM L-glutamine; 100 U/mL penicillin; and 100 µg/mL streptomycin sulfate. Store at 2-8ºC, protected from light for no longer than one month. Before use, warm in a 37ºC water bath.</p></dd><dt>4.3.</dt><dd id="ncipr54.lt31"><p class="no_top_margin">
|
||
<u>Complete Alpha MEM (to maintain NK92 cells)</u>
|
||
</p><p>The complete Alpha MEM medium should contain the following reagents: 2 mM L-glutamine, 0.2 mM inositol, 0.1 mM β-mercaptoethanol, 0.02 mM folic acid, 25 ng/mL recombinant IL-2, 10% horse serum, and 10% heat inactivated fetal bovine serum.</p></dd></dl></div><div id="ncipr54.s5"><h2 id="_ncipr54_s5_">5. Preparation of Study Samples</h2><p>This assay requires 3 mL of the nanoparticle at a concentration 10 × above the highest final concentration to be tested. Nanoparticles should be dissolved/resuspended in alpha-MEM medium. The concentration is selected based on the plasma concentration of the nanoparticle at the intended therapeutic dose. For the purpose of this protocol this concentration is called “theoretical plasma concentration”. Considerations for estimating theoretical plasma concentration were reviewed elsewhere [<a class="bk_pop" href="#ncipr54.ref3">3</a>] and are summarized in <a href="/books/NBK604916/box/ncipr54.box16/?report=objectonly" target="object" rid-ob="figobncipr54box16">Box 1</a> below.</p><p>The assay will evaluate 4 concentrations: 10X (or when feasible 100X, 30X or 5X) of the theoretical plasma concentration, theoretical plasma concentration and two 1: 5 serial dilutions of the theoretical plasma concentration. When the intended therapeutic concentration is unknown, the highest final concentration is 1 mg/mL or the highest reasonably achievable concentration.</p><p>For example, if the final theoretical plasma concentration to be tested is 0.2 mg/mL, then a stock of 20 mg/mL will be prepared and diluted 10-fold (2 mg/mL), followed by two 1: 5 serial dilutions (0.4 and 0.08 mg/mL). When 1.5 mL of each of these samples are combined in a T25 flask with 13.5 mL of cells, the final concentrations of nanoparticles are 0.008, 0.04, 0.2, and 2mg/mL. Each nanoparticle concentration is tested in duplicate. Additional 200 μL is required for cell free control.</p><div id="ncipr54.box16" class="box"><h3><span class="label">Box 1</span><span class="title">Example Calculation to Determine Nanoparticle Concentration for In Vitro Tests</span></h3><p>In this example, we assume a mouse dose of 123 mg/kg. Therefore, the scaled equivalent human dose would be:
|
||
<div class="pmc_disp_formula whole_rhythm clearfix" id="ncipr54.deq1"><div class="inline_block pmc_inline_block pmc_va_middle pmc_hide_overflow twelve_col"><math id="ncipr54.eq1" display="block"><mi>H</mi><mi>u</mi><mi>m</mi><mi>a</mi><mi>n</mi><mtext> </mtext><mi>d</mi><mi>o</mi><mi>s</mi><mi>e</mi><mo>=</mo><mfrac><mrow><mi>m</mi><mi>o</mi><mi>u</mi><mi>s</mi><mi>e</mi><mtext> </mtext><mi>d</mi><mi>o</mi><mi>s</mi><mi>e</mi></mrow><mrow><mn>12.3</mn></mrow></mfrac><mo>=</mo><mfrac><mrow><mn>123</mn><mtext> </mtext><mi>m</mi><mi>g</mi><mo>/</mo><mi>k</mi><mi>g</mi></mrow><mrow><mn>12.3</mn></mrow></mfrac><mo>=</mo><mn>10</mn><mtext> </mtext><mi>m</mi><mi>g</mi><mo>/</mo><mi>k</mi><mi>g</mi></math></div><div class="inline_block pmc_inline_block pmc_va_middle pmc_hide_overflow last bk_equ_label "><div><span class="nowrap"></span></div></div></div>
|
||
Blood volume constitutes approximately 8% of the body weight. Therefore, an average human of 70 kg body weight has approximately 5.6 L of blood. Assuming all the nanoparticle injected goes into the systemic circulation, this provides a rough approximation of the potential maximum nanoparticle concentration in blood, which is used as the in vitro test concentration.</p><div class="pmc_disp_formula whole_rhythm clearfix" id="ncipr54.deq2"><div class="inline_block pmc_inline_block pmc_va_middle pmc_hide_overflow twelve_col">
|
||
<math id="ncipr54.eq2" display="block"><mi>i</mi><mi>n</mi><mtext> </mtext><mi>v</mi><mi>i</mi><mi>t</mi><mi>r</mi><mi>o</mi><mtext> </mtext><mi>c</mi><mi>o</mi><mi>n</mi><mi>c</mi><mi>e</mi><mi>n</mi><mi>t</mi><mi>r</mi><mi>a</mi><mi>t</mi><mi>i</mi><mi>o</mi><msub><mi>n</mi><mrow><mi>h</mi><mi>u</mi><mi>m</mi><mi>a</mi><mi>n</mi><mtext> </mtext><mi>m</mi><mi>a</mi><mi>t</mi><mi>r</mi><mi>i</mi><mi>x</mi></mrow></msub><mo>=</mo><mfrac><mrow><mi>h</mi><mi>u</mi><mi>m</mi><mi>a</mi><mi>n</mi><mtext> </mtext><mi>d</mi><mi>o</mi><mi>s</mi><mi>e</mi></mrow><mrow><mi>h</mi><mi>u</mi><mi>m</mi><mi>a</mi><mi>n</mi><mtext> </mtext><mi>b</mi><mi>l</mi><mi>o</mi><mi>o</mi><mi>d</mi><mtext> </mtext><mi>v</mi><mi>o</mi><mi>l</mi><mi>u</mi><mi>m</mi><mi>e</mi></mrow></mfrac><mo>=</mo><mfrac><mrow><mn>70</mn><mtext> </mtext><mi>k</mi><mi>g</mi><mtext> </mtext><mo>∗</mo><mtext> </mtext><mn>10</mn><mtext> </mtext><mi>m</mi><mi>g</mi><mo>/</mo><mi>k</mi><mi>g</mi></mrow><mrow><mn>5.6</mn><mtext> </mtext><mi>L</mi></mrow></mfrac><mo>=</mo><mn>0.125</mn><mtext> </mtext><mi>m</mi><mi>g</mi><mo>/</mo><mi>m</mi><mi>L</mi></math>
|
||
</div><div class="inline_block pmc_inline_block pmc_va_middle pmc_hide_overflow last bk_equ_label "><div><span class="nowrap"></span></div></div></div></div></div><div id="ncipr54.s6"><h2 id="_ncipr54_s6_">6. Preparation of Effector and Target Cells</h2><dl id="ncipr54.l6" class="temp-labeled-list"><dt>6.1.</dt><dd id="ncipr54.lt32"><p class="no_top_margin">
|
||
<u>Preparation of NK92 Effector Cells</u>
|
||
</p><p>Grow cells in complete alpha-MEM medium. Split cells when the cell number approaches 1 × 10<sup>6</sup> cells/mL (i.e. approximately every 2-3 days). Do not allow cells to grow over 1 × 10<sup>6</sup> cells/mL. Cultures can be maintained by addition or replacement of medium. When replacing media, centrifuge cells at 130xg for 10 min, and resuspend the cell pellet in fresh medium at 2 × 10<sup>5</sup> to 3 × 10<sup>5</sup> viable cells/mL. Pipet the cells up and down on the back of the flask every 2-3 days to produce a single cell suspension. NK92 cells are extremely sensitive to overgrowth and media exhaustion. Replace with fresh medium every 2 to 3 days (depending on cell density).</p></dd><dt>6.2.</dt><dd id="ncipr54.lt33"><p class="no_top_margin">
|
||
<u>Preparation of HepG2 Target Cells</u>
|
||
</p><p>Grow cells in complete RPMI medium. Renew growth media twice a week. A subcultivation ratio of 1:4 or 1:6 is recommended. To split the cells, remove and discard culture medium; briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum (contains trypsin inhibitor). Then, add 2.0 to 3.0 mL of Trypsin-EDTA solution to the flask and observe cells under an inverted microscope until the cell layer is dispersed (usually within 2 to 5 minutes).</p><p>To avoid clumping, do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37ºC to facilitate dispersal. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture flasks and incubate cultures at 37ºC in 5% CO<sub>2</sub> environment.</p></dd></dl></div><div id="ncipr54.s7"><h2 id="_ncipr54_s7_">7. Experimental Procedure (This will require 3 days.)</h2><ul id="ncipr54.l7" class="simple-list"><li id="ncipr54.lt34" class="half_rhythm"><div><b><u>Day 1</u></b>
|
||
<dl id="ncipr54.l8" class="temp-labeled-list"><dt>7.1.</dt><dd id="ncipr54.lt35"><p class="no_top_margin">Adjust effector cell (NK92) number to 1 × 10<sup>6</sup> cells/mL using complete <b><u>alpha-MEM</u></b>. Prepare 10-15 mL of cell suspension for each sample and control (negative and vehicle).</p></dd><dt>7.2.</dt><dd id="ncipr54.lt36"><p class="no_top_margin">Treat NK92 cells with test nanoparticles, vehicle or negative control for 24 ± 0.5 hours. Perform treatment in T25 flask.</p></dd><dt>7.3.</dt><dd id="ncipr54.lt37"><p class="no_top_margin">Adjust target cell (HepG2) number to 0.5 × 10<sup>6</sup> cells/mL using complete <b><u>RPMI</u></b>.</p></dd><dt>7.4.</dt><dd id="ncipr54.lt38"><p class="no_top_margin">Plate 50 µL of media to all wells, attach plate to RT-CES and begin program. Following background measurement, plate 50 µL of HepG2 cells from <a href="#ncipr54.lt37">step 7.3</a> per each well in RT-CES plates except for nanoparticles only wells (please refer to the template in <a class="figpopup" href="/books/NBK604916/figure/ncipr54.fig1/?report=objectonly" target="object" rid-figpopup="figncipr54fig1" rid-ob="figobncipr54fig1">Figure 1</a> and remember that one needs 2 E-plates per each nanoparticle), attach to RT CES and start data acquisition. HepG2 cells are in culture for ~ 16 - 20 hr prior to addition of NK92 effector cells. Acquisition program is described in <a href="/books/NBK604916/table/ncipr54.tab1all/?report=objectonly" target="object" rid-ob="figobncipr54tab1all">Table 1</a>. Either version A or B can be used.</p></dd></dl></div></li><li id="ncipr54.lt39" class="half_rhythm"><div><b><u>Day 2</u></b>
|
||
<dl id="ncipr54.l9" class="temp-labeled-list"><dt>7.5.</dt><dd id="ncipr54.lt40"><p class="no_top_margin">Using Trypan Blue, determine viability of NK92 cells prepared in <a href="#ncipr54.lt36">step 7.2</a>.</p></dd><dt>7.6.</dt><dd id="ncipr54.lt41"><p class="no_top_margin">Concentrate NK92 cells from <a href="#ncipr54.lt40">step 7.5</a>. by centrifugation (5 min, 400xg); remove and discard supernatants, and reconstitute cells in each sample with fresh alpha-MEM media to the final concentration of 2.5 × 10<sup>6</sup> viable cells/mL using complete <b><u>alpha-MEM (without IL-2)</u></b>. This concentration will allow for an effector to target (E:T) ratio of 5:1. E:T ratio of 2.5:1 or 1.25:1 is also acceptable to use, and in this case the NK92 concentration should be adjusted to 1.25 × 10<sup>6</sup> or 0.625 × 10<sup>6</sup> viable cells/mL. If the viability of NK cells treated with negative control is ≥ 97%, proceed to the next step.</p><p><b>Note</b>: If nanoparticles were cytotoxic to NK92 cells and resulted in more than 50% cell death, it may not be possible to evaluate the cytotoxicity of NK92 cells treated with these nanoparticles due to a lack of the required number of effector cells.</p></dd><dt>7.7.</dt><dd id="ncipr54.lt42"><p class="no_top_margin">Pause RT-CES data acquisition program; remove RT-CES plates from the instrument and add 100 µL of NK cells from <a href="#ncipr54.lt41">step 7.6</a> to designated wells on RT-CES plate. An example of a template is provided in <a class="figpopup" href="/books/NBK604916/figure/ncipr54.fig1/?report=objectonly" target="object" rid-figpopup="figncipr54fig1" rid-ob="figobncipr54fig1">Figure 1</a>. Prepare two E-plates for each nanoparticle.</p></dd><dt>7.8.</dt><dd id="ncipr54.lt43"><p class="no_top_margin">Return RT-CES plates containing NK92 effector cells treated with either nanoparticle or negative control and target (HepG2) cells to the instrument and resume data acquisition for another 24 hr.</p></dd></dl></div></li><li id="ncipr54.lt44" class="half_rhythm"><div><b><u>Day 3</u></b>
|
||
<dl id="ncipr54.l10" class="temp-labeled-list"><dt>7.9.</dt><dd id="ncipr54.lt45"><p class="no_top_margin">Stop acquisition program on the RT-CES instrument and analyze the data.</p></dd></dl></div></li></ul><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figncipr54tab1all"><a href="/books/NBK604916/table/ncipr54.tab1all/?report=objectonly" target="object" title="Table 1" class="img_link icnblk_img" rid-ob="figobncipr54tab1all"><img class="small-thumb" src="/corehtml/pmc/css/bookshelf/2.26/img/table-icon.gif" alt="Table Icon" /></a><div class="icnblk_cntnt"><h4 id="ncipr54.tab1all"><a href="/books/NBK604916/table/ncipr54.tab1all/?report=objectonly" target="object" rid-ob="figobncipr54tab1all">Table 1</a></h4><p class="float-caption no_bottom_margin">RT-CES Acquisition Protocols. </p></div></div><div class="iconblock whole_rhythm clearfix ten_col fig" id="figncipr54fig1" co-legend-rid="figlgndncipr54fig1"><a href="/books/NBK604916/figure/ncipr54.fig1/?report=objectonly" target="object" title="Figure 1" class="img_link icnblk_img figpopup" rid-figpopup="figncipr54fig1" rid-ob="figobncipr54fig1"><img class="small-thumb" src="/books/NBK604916/bin/ncipr54f1.gif" src-large="/books/NBK604916/bin/ncipr54f1.jpg" alt="Figure 1. Example of RT-CES Plate Template." /></a><div class="icnblk_cntnt" id="figlgndncipr54fig1"><h4 id="ncipr54.fig1"><a href="/books/NBK604916/figure/ncipr54.fig1/?report=objectonly" target="object" rid-ob="figobncipr54fig1">Figure 1</a></h4><p class="float-caption no_bottom_margin">Example of RT-CES Plate Template. Nanoparticles are tested at 4 concentrations. Testing of the “nanoparticles only” sample is recommended to identify potential nanoparticle interference with the RT-CES instrument acquisition system. During <a href="/books/NBK604916/figure/ncipr54.fig1/?report=objectonly" target="object" rid-ob="figobncipr54fig1">(more...)</a></p></div></div></div><div id="ncipr54.s8"><h2 id="_ncipr54_s8_">8. Calculations</h2><p>Example of cell index plot is shown in <a class="figpopup" href="/books/NBK604916/figure/ncipr54.fig2/?report=objectonly" target="object" rid-figpopup="figncipr54fig2" rid-ob="figobncipr54fig2">Figure 2</a>. Overview of the instrument and how it functions is shown in <a class="figpopup" href="/books/NBK604916/figure/ncipr54.fig3/?report=objectonly" target="object" rid-figpopup="figncipr54fig3" rid-ob="figobncipr54fig3">Figure 3</a>. Cell index data for each test sample and control is used to calculate area under the curve (AUC). The AUC data from each control and test sample is used to calculate percent cytotoxicity and percent coefficient of variation (CV). The %CV is used to control precision and is calculated according to the following formula:
|
||
<div class="pmc_disp_formula whole_rhythm clearfix" id="ncipr54.deq3"><div class="inline_block pmc_inline_block pmc_va_middle pmc_hide_overflow twelve_col"><math id="ncipr54.eq3" display="block"><mrow><mi>%</mi><mi>C</mi><mi>V</mi><mo>=</mo><mfrac><mrow><mi>s</mi><mi>t</mi><mi>a</mi><mi>n</mi><mi>d</mi><mi>a</mi><mi>r</mi><mi>d</mi><mtext> </mtext><mi>d</mi><mi>e</mi><mi>v</mi><mi>i</mi><mi>a</mi><mi>t</mi><mi>i</mi><mi>o</mi><mi>n</mi></mrow><mrow><mi>m</mi><mi>e</mi><mi>a</mi><mi>n</mi></mrow></mfrac><mo>∗</mo><mn>100</mn><mi>%</mi></mrow></math></div><div class="inline_block pmc_inline_block pmc_va_middle pmc_hide_overflow last bk_equ_label "><div><span class="nowrap"></span></div></div></div>
|
||
<div class="pmc_disp_formula whole_rhythm clearfix" id="ncipr54.deq4"><div class="inline_block pmc_inline_block pmc_va_middle pmc_hide_overflow twelve_col"><math id="ncipr54.eq4" display="block"><mrow><mi>%</mi><mi>C</mi><mi>y</mi><mi>t</mi><mi>o</mi><mi>t</mi><mi>o</mi><mi>x</mi><mi>i</mi><mi>c</mi><mi>i</mi><mi>t</mi><mi>y</mi><mo>=</mo><mfrac><mrow><mi>A</mi><mi>U</mi><msub><mi>C</mi><mrow><mi>b</mi><mi>a</mi><mi>s</mi><mi>e</mi><mi>l</mi><mi>i</mi><mi>n</mi><mi>e</mi></mrow></msub><mo>−</mo><mi>A</mi><mi>U</mi><msub><mi>C</mi><mrow><mi>p</mi><mi>a</mi><mi>r</mi><mi>t</mi><mi>i</mi><mi>c</mi><mi>l</mi><mi>e</mi><mi>s</mi></mrow></msub></mrow><mrow><mi>A</mi><mi>U</mi><msub><mi>C</mi><mrow><mi>b</mi><mi>a</mi><mi>s</mi><mi>e</mi><mi>l</mi><mi>i</mi><mi>n</mi><mi>e</mi></mrow></msub></mrow></mfrac><mo>∗</mo><mn>100</mn><mi>%</mi></mrow></math></div><div class="inline_block pmc_inline_block pmc_va_middle pmc_hide_overflow last bk_equ_label "><div><span class="nowrap"></span></div></div></div>
|
||
where AUC is area under the curve determined for HepG2 growth from the time of addition of NK2 effector cells to the time when the data acquisition was stopped (i.e. 24 ± 0.5 hr later) and normalized to the number of cells plated in each individual well.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figncipr54fig2" co-legend-rid="figlgndncipr54fig2"><a href="/books/NBK604916/figure/ncipr54.fig2/?report=objectonly" target="object" title="Figure 2" class="img_link icnblk_img figpopup" rid-figpopup="figncipr54fig2" rid-ob="figobncipr54fig2"><img class="small-thumb" src="/books/NBK604916/bin/ncipr54f2.gif" src-large="/books/NBK604916/bin/ncipr54f2.jpg" alt="Graph of raw data. Time displayed on x-axis. Cell index displayed on y-axis." /></a><div class="icnblk_cntnt" id="figlgndncipr54fig2"><h4 id="ncipr54.fig2"><a href="/books/NBK604916/figure/ncipr54.fig2/?report=objectonly" target="object" rid-ob="figobncipr54fig2">Figure 2</a></h4><p class="float-caption no_bottom_margin">Example of raw data demonstrating changes in HepG2 cell index after co-incubation with NK92 cells. </p></div></div><div class="iconblock whole_rhythm clearfix ten_col fig" id="figncipr54fig3" co-legend-rid="figlgndncipr54fig3"><a href="/books/NBK604916/figure/ncipr54.fig3/?report=objectonly" target="object" title="Figure 3" class="img_link icnblk_img figpopup" rid-figpopup="figncipr54fig3" rid-ob="figobncipr54fig3"><img class="small-thumb" src="/books/NBK604916/bin/ncipr54f3.gif" src-large="/books/NBK604916/bin/ncipr54f3.jpg" alt="Top left: Photograph of the RT-CES instrument and computer. Top right: Photograph of a 96-well plate. Bottom left: Cartoon depiction showing how electric impedance increases with increasing number of cells attached. Bottom right: Graph showing increased impedance with increased time. Also, microscope picture of well containing cells for each time point." /></a><div class="icnblk_cntnt" id="figlgndncipr54fig3"><h4 id="ncipr54.fig3"><a href="/books/NBK604916/figure/ncipr54.fig3/?report=objectonly" target="object" rid-ob="figobncipr54fig3">Figure 3</a></h4><p class="float-caption no_bottom_margin">Overview of RT-CES Instrumentation. The images above are reproduced from the reference 4, also named on the image, and are used for the informational purposes only. </p></div></div></div><div id="ncipr54.s9"><h2 id="_ncipr54_s9_">9. Acceptance Criteria</h2><dl id="ncipr54.l11" class="temp-labeled-list"><dt>9.1.</dt><dd id="ncipr54.lt46"><p class="no_top_margin">CV of test samples and control should be within 25%.</p></dd><dt>9.2.</dt><dd id="ncipr54.lt47"><p class="no_top_margin">Assay is acceptable if percent cytotoxicity in negative control is ≥ 30%.</p></dd></dl></div><div id="ncipr54.rl.r1"><h2 id="_ncipr54_rl_r1_">10. References</h2><dl class="temp-labeled-list"><dt>1.</dt><dd><div class="bk_ref" id="ncipr54.ref1">Abassi, Y.; Zhu, J.
|
||
Label-free assay for NK cell mediated cytolysis on the RT-CES system. RT-CES applications. ACEA Biosciences;<a href="http://www.aceabio.com/" ref="pagearea=cite-ref&targetsite=external&targetcat=link&targettype=uri">www<wbr style="display:inline-block"></wbr>.aceabio.com</a>.</div></dd><dt>2.</dt><dd><div class="bk_ref" id="ncipr54.ref2">Zhu, J.; Wang, X.; Xu, X.; Abassi, Y.A.
|
||
Dynamic and label-free monitoring of natural killer cell cytotoxic activity using electornic cell sensor arrays. <em>Journal of Immunological Methods</em> (2006) 309: 25–33
|
||
[<a href="https://pubmed.ncbi.nlm.nih.gov/16423365" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 16423365</span></a>]</div></dd><dt>3.</dt><dd><div class="bk_ref" id="ncipr54.ref3">Dobrovolskaia
|
||
MA, McNeil
|
||
SE. Understanding the correlation between in vitro and in vivo immunotoxicity tests for nanomedicines. J Control Release. 2013
|
||
Dec10; 172(2): 456–66
|
||
[<a href="/pmc/articles/PMC5831149/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC5831149</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/23742883" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 23742883</span></a>]</div></dd><dt>4.</dt><dd><div class="bk_ref" id="ncipr54.ref4">Solly
|
||
K, Wang
|
||
X, Xu
|
||
X, Strulovici
|
||
B, Zheng
|
||
W. Application of real-time cell electronic sensing (RT-CES) technology to cell-based assays. Assay Drug Dev Technol. 2004;2(4):363–372. doi:10.1089/adt.2004.2.363
|
||
[<a href="https://pubmed.ncbi.nlm.nih.gov/15357917" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 15357917</span></a>] [<a href="http://dx.crossref.org/10.1089/adt.2004.2.363" ref="pagearea=cite-ref&targetsite=external&targetcat=link&targettype=uri">CrossRef</a>]</div></dd></dl></div><div id="ncipr54.s10"><h2 id="_ncipr54_s10_">11. Abbreviations</h2><dl><dt id="ncipr54.abb_DL1_DI1">AUC</dt><dd><p>area under the curve</p></dd><dt id="ncipr54.abb_DL1_DI2">CV</dt><dd><p>coefficient of variation</p></dd><dt id="ncipr54.abb_DL1_DI3">E:T</dt><dd><p>effector to target cell ratio</p></dd><dt id="ncipr54.abb_DL1_DI4">FBS</dt><dd><p>fetal bovine serum</p></dd><dt id="ncipr54.abb_DL1_DI5">HepG2</dt><dd><p>human hepatocarcinoma cells</p></dd><dt id="ncipr54.abb_DL1_DI6">IL</dt><dd><p>interleukin</p></dd><dt id="ncipr54.abb_DL1_DI7">MEM</dt><dd><p>minimal essential medium</p></dd><dt id="ncipr54.abb_DL1_DI8">NK</dt><dd><p>natural killer</p></dd><dt id="ncipr54.abb_DL1_DI9">PAMAM</dt><dd><p>polyamidoamine</p></dd><dt id="ncipr54.abb_DL1_DI10">PBS</dt><dd><p>phosphate buffered saline</p></dd><dt id="ncipr54.abb_DL1_DI11">RT-CES</dt><dd><p>real-time cell electronic sensing</p></dd><dt id="ncipr54.abb_DL1_DI12">SD</dt><dd><p>standard deviation</p></dd></dl></div><div><dl class="temp-labeled-list small"><dt></dt><dd><div><p class="no_top_margin"><p>This protocol assumes an intermediate level of scientific competency with regard to techniques, instrumentation, and safety procedures. Rudimentary assay details have been omitted for the sake of brevity.</p></p></div></dd><dt>*</dt><dd><div id="ncipr54.fn1"><p class="no_top_margin">address correspondence to: <a href="mailto:dev@null" data-email="vog.hin.liam@aniram" class="oemail">vog.hin.liam@aniram</a></p></div></dd><dt></dt><dd><div><p class="no_top_margin"><div>
|
||
<span class="mixed-citation" id="ncipr54.suggestedcitation">Potter TM, Cedrone E, Neun BW, Dobrovolskaia MA, NCL Method ITA-11: Measurement of Nanoparticle Effects on Cytotoxic Activity of NK Cells by Label-Free RT-CES System. <a href="https://ncl.cancer.gov/resources/assay-cascade-protocols" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">https://ncl.cancer.gov/resources/assay-cascade-protocols</a> DOI: <a href="http://dx.crossref.org/10.17917/GMXG-BH29" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">10.17917/GMXG-BH29</a></span>
|
||
</div></p></div></dd></dl></div><div id="bk_toc_contnr"></div></div></div>
|
||
<div class="post-content"><div><div class="half_rhythm"><a href="/books/about/copyright/">Copyright Notice</a></div><div class="small"><span class="label">Bookshelf ID: NBK604916</span><span class="label">PMID: <a href="https://pubmed.ncbi.nlm.nih.gov/39013070" title="PubMed record of this page" ref="pagearea=meta&targetsite=entrez&targetcat=link&targettype=pubmed">39013070</a></span>DOI: <a href="http://dx.crossref.org/10.17917/GMXG-BH29" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">10.17917/GMXG-BH29</a></div><div style="margin-top:2em" class="bk_noprnt"><a class="bk_cntns" href="/books/n/nciprotocols/">Contents</a><div class="pagination bk_noprnt"><a class="active page_link prev" href="/books/n/nciprotocols/ncipr55/" title="Previous page in this title">< Prev</a><a class="active page_link next" href="/books/n/nciprotocols/ncipr49/" title="Next page in this title">Next ></a></div></div></div></div>
|
||
|
||
</div>
|
||
|
||
<!-- Custom content below content -->
|
||
<div class="col4">
|
||
|
||
</div>
|
||
|
||
|
||
<!-- Book content -->
|
||
|
||
<!-- Custom contetnt below bottom nav -->
|
||
<div class="col5">
|
||
|
||
</div>
|
||
</div>
|
||
|
||
<div id="rightcolumn" class="four_col col last">
|
||
<!-- Custom content above discovery portlets -->
|
||
<div class="col6">
|
||
<div id="ncbi_share_book"><a href="#" class="ncbi_share" data-ncbi_share_config="popup:false,shorten:true" ref="id=NBK604916&db=books">Share</a></div>
|
||
|
||
</div>
|
||
<div xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance"></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Views</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="PDF_download" id="Shutter"></a></div><div class="portlet_content"><ul xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="simple-list"><li><a href="/books/NBK604916/?report=reader">PubReader</a></li><li><a href="/books/NBK604916/?report=printable">Print View</a></li><li><a data-jig="ncbidialog" href="#_ncbi_dlg_citbx_NBK604916" data-jigconfig="width:400,modal:true">Cite this Page</a><div id="_ncbi_dlg_citbx_NBK604916" style="display:none" title="Cite this Page"><div class="bk_tt">Potter TM, Cedrone E, Neun BW, et al. Measurement of Nanoparticle Effects on Cytotoxic Activity of NK Cells by Label-Free RT-CES System: Version 3. 2020 Jun. In: National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols [Internet]. Bethesda (MD): National Cancer Institute (US); 2005 May 1-. NCL Method ITA-11.<span class="bk_cite_avail"></span> doi: 10.17917/GMXG-BH29</div></div></li><li><a href="/books/NBK604916/pdf/Bookshelf_NBK604916.pdf">PDF version of this page</a> (562K)</li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>In this Page</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="page-toc" id="Shutter"></a></div><div class="portlet_content"><ul xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="simple-list"><li><a href="#ncipr54.s1" ref="log$=inpage&link_id=inpage">Introduction</a></li><li><a href="#ncipr54.s2" ref="log$=inpage&link_id=inpage">Principles</a></li><li><a href="#ncipr54.s3" ref="log$=inpage&link_id=inpage">Reagents, Materials, and Equipment</a></li><li><a href="#ncipr54.s4" ref="log$=inpage&link_id=inpage">Reagent Preparation</a></li><li><a href="#ncipr54.s5" ref="log$=inpage&link_id=inpage">Preparation of Study Samples</a></li><li><a href="#ncipr54.s6" ref="log$=inpage&link_id=inpage">Preparation of Effector and Target Cells</a></li><li><a href="#ncipr54.s7" ref="log$=inpage&link_id=inpage">Experimental Procedure (This will require 3 days.)</a></li><li><a href="#ncipr54.s8" ref="log$=inpage&link_id=inpage">Calculations</a></li><li><a href="#ncipr54.s9" ref="log$=inpage&link_id=inpage">Acceptance Criteria</a></li><li><a href="#ncipr54.rl.r1" ref="log$=inpage&link_id=inpage">References</a></li><li><a href="#ncipr54.s10" ref="log$=inpage&link_id=inpage">Abbreviations</a></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Related information</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="discovery_db_links" id="Shutter"></a></div><div class="portlet_content"><ul><li class="brieflinkpopper"><a class="brieflinkpopperctrl" href="/books/?Db=pmc&DbFrom=books&Cmd=Link&LinkName=books_pmc_refs&IdsFromResult=5640950" ref="log$=recordlinks">PMC</a><div class="brieflinkpop offscreen_noflow">PubMed Central citations</div></li><li class="brieflinkpopper"><a class="brieflinkpopperctrl" href="/books/?Db=pubmed&DbFrom=books&Cmd=Link&LinkName=books_pubmed_refs&IdsFromResult=5640950" ref="log$=recordlinks">PubMed</a><div class="brieflinkpop offscreen_noflow">Links to PubMed</div></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Similar articles in PubMed</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="PBooksDiscovery_RA" id="Shutter"></a></div><div class="portlet_content"><ul><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/29882143" ref="ordinalpos=1&linkpos=1&log$=relatedarticles&logdbfrom=pubmed">Natural Killer (NK) Cell Assays in Immunotoxicity Testing.</a><span class="source">[Methods Mol Biol. 2018]</span><div class="brieflinkpop offscreen_noflow">Natural Killer (NK) Cell Assays in Immunotoxicity Testing.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Li Q. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Methods Mol Biol. 2018; 1803:231-241. </em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/17507786" ref="ordinalpos=1&linkpos=2&log$=relatedreviews&logdbfrom=pubmed"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> New mechanism of organophosphorus pesticide-induced immunotoxicity.</a><span class="source">[J Nippon Med Sch. 2007]</span><div class="brieflinkpop offscreen_noflow"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> New mechanism of organophosphorus pesticide-induced immunotoxicity.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Li Q. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">J Nippon Med Sch. 2007 Apr; 74(2):92-105. </em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/23632890" ref="ordinalpos=1&linkpos=3&log$=relatedarticles&logdbfrom=pubmed">LAMP1/CD107a is required for efficient perforin delivery to lytic granules and NK-cell cytotoxicity.</a><span class="source">[Blood. 2013]</span><div class="brieflinkpop offscreen_noflow">LAMP1/CD107a is required for efficient perforin delivery to lytic granules and NK-cell cytotoxicity.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Krzewski K, Gil-Krzewska A, Nguyen V, Peruzzi G, Coligan JE. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Blood. 2013 Jun 6; 121(23):4672-83. Epub 2013 Apr 30.</em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/35202434" ref="ordinalpos=1&linkpos=4&log$=relatedarticles&logdbfrom=pubmed">Natural killer cells kill extracellular Pseudomonas aeruginosa using contact-dependent release of granzymes B and H.</a><span class="source">[PLoS Pathog. 2022]</span><div class="brieflinkpop offscreen_noflow">Natural killer cells kill extracellular Pseudomonas aeruginosa using contact-dependent release of granzymes B and H.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Feehan DD, Jamil K, Polyak MJ, Ogbomo H, Hasell M, Li SS, Xiang RF, Parkins M, Trapani JA, Harrison JJ, et al. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">PLoS Pathog. 2022 Feb; 18(2):e1010325. Epub 2022 Feb 24.</em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/8011296" ref="ordinalpos=1&linkpos=5&log$=relatedreviews&logdbfrom=pubmed"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> The binding and lysis of target cells by cytotoxic lymphocytes: molecular and cellular aspects.</a><span class="source">[Annu Rev Immunol. 1994]</span><div class="brieflinkpop offscreen_noflow"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> The binding and lysis of target cells by cytotoxic lymphocytes: molecular and cellular aspects.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Berke G. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Annu Rev Immunol. 1994; 12:735-73. </em></div></div></li></ul><a class="seemore" href="/sites/entrez?db=pubmed&cmd=link&linkname=pubmed_pubmed_reviews&uid=39013070" ref="ordinalpos=1&log$=relatedreviews_seeall&logdbfrom=pubmed">See reviews...</a><a class="seemore" href="/sites/entrez?db=pubmed&cmd=link&linkname=pubmed_pubmed&uid=39013070" ref="ordinalpos=1&log$=relatedarticles_seeall&logdbfrom=pubmed">See all...</a></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Recent Activity</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="recent_activity" id="Shutter"></a></div><div class="portlet_content"><div xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" id="HTDisplay" class=""><div class="action"><a href="javascript:historyDisplayState('ClearHT')">Clear</a><a href="javascript:historyDisplayState('HTOff')" class="HTOn">Turn Off</a><a href="javascript:historyDisplayState('HTOn')" class="HTOff">Turn On</a></div><ul id="activity"><li class="ra_rcd ralinkpopper two_line"><a class="htb ralinkpopperctrl" ref="log$=activity&linkpos=1" href="/portal/utils/pageresolver.fcgi?recordid=67d8488767c23b31e0154991">Measurement of Nanoparticle Effects on Cytotoxic Activity of NK Cells by Label-F...</a><div class="ralinkpop offscreen_noflow">Measurement of Nanoparticle Effects on Cytotoxic Activity of NK Cells by Label-Free RT-CES System - National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols<div class="brieflinkpopdesc"></div></div><div class="tertiary"></div></li><li class="ra_rcd ralinkpopper two_line"><a class="htb ralinkpopperctrl" ref="log$=activity&linkpos=2" href="/portal/utils/pageresolver.fcgi?recordid=67d848862f30673f7b5d9fc3">Figure 3, Overview of RT-CES Instrumentation - National Cancer Institute’s Nanot...</a><div class="ralinkpop offscreen_noflow">Figure 3, Overview of RT-CES Instrumentation - National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols<div class="brieflinkpopdesc"></div></div><div class="tertiary"></div></li><li class="ra_rcd ralinkpopper two_line"><a class="htb ralinkpopperctrl" ref="log$=activity&linkpos=3" href="/portal/utils/pageresolver.fcgi?recordid=67d848852f30673f7b5d9821">Figure 2, Example of raw data demonstrating changes in HepG2 cell index after co...</a><div class="ralinkpop offscreen_noflow">Figure 2, Example of raw data demonstrating changes in HepG2 cell index after co-incubation with NK92 cells - National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols<div class="brieflinkpopdesc"></div></div><div class="tertiary"></div></li><li class="ra_rcd ralinkpopper two_line"><a class="htb ralinkpopperctrl" ref="log$=activity&linkpos=4" href="/portal/utils/pageresolver.fcgi?recordid=67d8488384f3725e5986bbd2">Table 1, RT-CES Acquisition Protocols - National Cancer Institute’s Nanotechnolo...</a><div class="ralinkpop offscreen_noflow">Table 1, RT-CES Acquisition Protocols - National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols<div class="brieflinkpopdesc"></div></div><div class="tertiary"></div></li><li class="ra_rcd ralinkpopper two_line"><a class="htb ralinkpopperctrl" ref="log$=activity&linkpos=5" href="/portal/utils/pageresolver.fcgi?recordid=67d84881cde49f3df74b79a6">Figure 1, Example of RT-CES Plate Template - National Cancer Institute’s Nanotec...</a><div class="ralinkpop offscreen_noflow">Figure 1, Example of RT-CES Plate Template - National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols<div class="brieflinkpopdesc"></div></div><div class="tertiary"></div></li></ul><p class="HTOn">Your browsing activity is empty.</p><p class="HTOff">Activity recording is turned off.</p><p id="turnOn" class="HTOff"><a href="javascript:historyDisplayState('HTOn')">Turn recording back on</a></p><a class="seemore" href="/sites/myncbi/recentactivity">See more...</a></div></div></div>
|
||
|
||
<!-- Custom content below discovery portlets -->
|
||
<div class="col7">
|
||
|
||
</div>
|
||
</div>
|
||
</div>
|
||
|
||
<!-- Custom content after all -->
|
||
<div class="col8">
|
||
|
||
</div>
|
||
<div class="col9">
|
||
|
||
</div>
|
||
|
||
<script type="text/javascript" src="/corehtml/pmc/js/jquery.scrollTo-1.4.2.js"></script>
|
||
<script type="text/javascript">
|
||
(function($){
|
||
$('.skiplink').each(function(i, item){
|
||
var href = $($(item).attr('href'));
|
||
href.attr('tabindex', '-1').addClass('skiptarget'); // ensure the target can receive focus
|
||
$(item).on('click', function(event){
|
||
event.preventDefault();
|
||
$.scrollTo(href, 0, {
|
||
onAfter: function(){
|
||
href.focus();
|
||
}
|
||
});
|
||
});
|
||
});
|
||
})(jQuery);
|
||
</script>
|
||
</div>
|
||
<div class="bottom">
|
||
|
||
<div id="NCBIFooter_dynamic">
|
||
<!--<component id="Breadcrumbs" label="breadcrumbs"/>
|
||
<component id="Breadcrumbs" label="helpdesk"/>-->
|
||
|
||
</div>
|
||
|
||
<div class="footer" id="footer">
|
||
<section class="icon-section">
|
||
<div id="icon-section-header" class="icon-section_header">Follow NCBI</div>
|
||
<div class="grid-container container">
|
||
<div class="icon-section_container">
|
||
<a class="footer-icon" id="footer_twitter" href="https://twitter.com/ncbi" aria-label="Twitter"><svg xmlns="http://www.w3.org/2000/svg" data-name="Layer 1" viewBox="0 0 300 300">
|
||
<defs>
|
||
<style>
|
||
.cls-11 {
|
||
fill: #737373;
|
||
}
|
||
</style>
|
||
</defs>
|
||
<title>Twitter</title>
|
||
<path class="cls-11" d="M250.11,105.48c-7,3.14-13,3.25-19.27.14,8.12-4.86,8.49-8.27,11.43-17.46a78.8,78.8,0,0,1-25,9.55,39.35,39.35,0,0,0-67,35.85,111.6,111.6,0,0,1-81-41.08A39.37,39.37,0,0,0,81.47,145a39.08,39.08,0,0,1-17.8-4.92c0,.17,0,.33,0,.5a39.32,39.32,0,0,0,31.53,38.54,39.26,39.26,0,0,1-17.75.68,39.37,39.37,0,0,0,36.72,27.3A79.07,79.07,0,0,1,56,223.34,111.31,111.31,0,0,0,116.22,241c72.3,0,111.83-59.9,111.83-111.84,0-1.71,0-3.4-.1-5.09C235.62,118.54,244.84,113.37,250.11,105.48Z">
|
||
</path>
|
||
</svg></a>
|
||
<a class="footer-icon" id="footer_facebook" href="https://www.facebook.com/ncbi.nlm" aria-label="Facebook"><svg xmlns="http://www.w3.org/2000/svg" data-name="Layer 1" viewBox="0 0 300 300">
|
||
<title>Facebook</title>
|
||
<path class="cls-11" d="M210.5,115.12H171.74V97.82c0-8.14,5.39-10,9.19-10h27.14V52l-39.32-.12c-35.66,0-42.42,26.68-42.42,43.77v19.48H99.09v36.32h27.24v109h45.41v-109h35Z">
|
||
</path>
|
||
</svg></a>
|
||
<a class="footer-icon" id="footer_linkedin" href="https://www.linkedin.com/company/ncbinlm" aria-label="LinkedIn"><svg xmlns="http://www.w3.org/2000/svg" data-name="Layer 1" viewBox="0 0 300 300">
|
||
<title>LinkedIn</title>
|
||
<path class="cls-11" d="M101.64,243.37H57.79v-114h43.85Zm-22-131.54h-.26c-13.25,0-21.82-10.36-21.82-21.76,0-11.65,8.84-21.15,22.33-21.15S101.7,78.72,102,90.38C102,101.77,93.4,111.83,79.63,111.83Zm100.93,52.61A17.54,17.54,0,0,0,163,182v61.39H119.18s.51-105.23,0-114H163v13a54.33,54.33,0,0,1,34.54-12.66c26,0,44.39,18.8,44.39,55.29v58.35H198.1V182A17.54,17.54,0,0,0,180.56,164.44Z">
|
||
</path>
|
||
</svg></a>
|
||
<a class="footer-icon" id="footer_github" href="https://github.com/ncbi" aria-label="GitHub"><svg xmlns="http://www.w3.org/2000/svg" data-name="Layer 1" viewBox="0 0 300 300">
|
||
<defs>
|
||
<style>
|
||
.cls-11,
|
||
.cls-12 {
|
||
fill: #737373;
|
||
}
|
||
|
||
.cls-11 {
|
||
fill-rule: evenodd;
|
||
}
|
||
</style>
|
||
</defs>
|
||
<title>GitHub</title>
|
||
<path class="cls-11" d="M151.36,47.28a105.76,105.76,0,0,0-33.43,206.1c5.28,1,7.22-2.3,7.22-5.09,0-2.52-.09-10.85-.14-19.69-29.42,6.4-35.63-12.48-35.63-12.48-4.81-12.22-11.74-15.47-11.74-15.47-9.59-6.56.73-6.43.73-6.43,10.61.75,16.21,10.9,16.21,10.9,9.43,16.17,24.73,11.49,30.77,8.79,1-6.83,3.69-11.5,6.71-14.14C108.57,197.1,83.88,188,83.88,147.51a40.92,40.92,0,0,1,10.9-28.39c-1.1-2.66-4.72-13.42,1-28,0,0,8.88-2.84,29.09,10.84a100.26,100.26,0,0,1,53,0C198,88.3,206.9,91.14,206.9,91.14c5.76,14.56,2.14,25.32,1,28a40.87,40.87,0,0,1,10.89,28.39c0,40.62-24.74,49.56-48.29,52.18,3.79,3.28,7.17,9.71,7.17,19.58,0,14.15-.12,25.54-.12,29,0,2.82,1.9,6.11,7.26,5.07A105.76,105.76,0,0,0,151.36,47.28Z">
|
||
</path>
|
||
<path class="cls-12" d="M85.66,199.12c-.23.52-1.06.68-1.81.32s-1.2-1.06-.95-1.59,1.06-.69,1.82-.33,1.21,1.07.94,1.6Zm-1.3-1">
|
||
</path>
|
||
<path class="cls-12" d="M90,203.89c-.51.47-1.49.25-2.16-.49a1.61,1.61,0,0,1-.31-2.19c.52-.47,1.47-.25,2.17.49s.82,1.72.3,2.19Zm-1-1.08">
|
||
</path>
|
||
<path class="cls-12" d="M94.12,210c-.65.46-1.71,0-2.37-.91s-.64-2.07,0-2.52,1.7,0,2.36.89.65,2.08,0,2.54Zm0,0"></path>
|
||
<path class="cls-12" d="M99.83,215.87c-.58.64-1.82.47-2.72-.41s-1.18-2.06-.6-2.7,1.83-.46,2.74.41,1.2,2.07.58,2.7Zm0,0">
|
||
</path>
|
||
<path class="cls-12" d="M107.71,219.29c-.26.82-1.45,1.2-2.64.85s-2-1.34-1.74-2.17,1.44-1.23,2.65-.85,2,1.32,1.73,2.17Zm0,0">
|
||
</path>
|
||
<path class="cls-12" d="M116.36,219.92c0,.87-1,1.59-2.24,1.61s-2.29-.68-2.3-1.54,1-1.59,2.26-1.61,2.28.67,2.28,1.54Zm0,0">
|
||
</path>
|
||
<path class="cls-12" d="M124.42,218.55c.15.85-.73,1.72-2,1.95s-2.37-.3-2.52-1.14.73-1.75,2-2,2.37.29,2.53,1.16Zm0,0"></path>
|
||
</svg></a>
|
||
<a class="footer-icon" id="footer_blog" href="https://ncbiinsights.ncbi.nlm.nih.gov/" aria-label="Blog">
|
||
<svg xmlns="http://www.w3.org/2000/svg" id="Layer_1" data-name="Layer 1" viewBox="0 0 40 40">
|
||
<defs><style>.cls-1{fill:#737373;}</style></defs>
|
||
<title>NCBI Insights Blog</title>
|
||
<path class="cls-1" d="M14,30a4,4,0,1,1-4-4,4,4,0,0,1,4,4Zm11,3A19,19,0,0,0,7.05,15a1,1,0,0,0-1,1v3a1,1,0,0,0,.93,1A14,14,0,0,1,20,33.07,1,1,0,0,0,21,34h3a1,1,0,0,0,1-1Zm9,0A28,28,0,0,0,7,6,1,1,0,0,0,6,7v3a1,1,0,0,0,1,1A23,23,0,0,1,29,33a1,1,0,0,0,1,1h3A1,1,0,0,0,34,33Z"></path>
|
||
</svg>
|
||
</a>
|
||
</div>
|
||
</div>
|
||
</section>
|
||
|
||
<section class="container-fluid bg-primary">
|
||
<div class="container pt-5">
|
||
<div class="row mt-3">
|
||
<div class="col-lg-3 col-12">
|
||
<p><a class="text-white" href="https://www.nlm.nih.gov/socialmedia/index.html">Connect with NLM</a></p>
|
||
<ul class="list-inline social_media">
|
||
<li class="list-inline-item"><a href="https://twitter.com/NLM_NIH" aria-label="Twitter" target="_blank" rel="noopener noreferrer"><svg xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink" version="1.1" x="0px" y="0px" viewBox="0 0 249 249" style="enable-background:new 0 0 249 249;" xml:space="preserve">
|
||
<style type="text/css">
|
||
.st20 {
|
||
fill: #FFFFFF;
|
||
}
|
||
|
||
.st30 {
|
||
fill: none;
|
||
stroke: #FFFFFF;
|
||
stroke-width: 8;
|
||
stroke-miterlimit: 10;
|
||
}
|
||
</style>
|
||
<title>Twitter</title>
|
||
<g>
|
||
<g>
|
||
<g>
|
||
<path class="st20" d="M192.9,88.1c-5,2.2-9.2,2.3-13.6,0.1c5.7-3.4,6-5.8,8.1-12.3c-5.4,3.2-11.4,5.5-17.6,6.7 c-10.5-11.2-28.1-11.7-39.2-1.2c-7.2,6.8-10.2,16.9-8,26.5c-22.3-1.1-43.1-11.7-57.2-29C58,91.6,61.8,107.9,74,116 c-4.4-0.1-8.7-1.3-12.6-3.4c0,0.1,0,0.2,0,0.4c0,13.2,9.3,24.6,22.3,27.2c-4.1,1.1-8.4,1.3-12.5,0.5c3.6,11.3,14,19,25.9,19.3 c-11.6,9.1-26.4,13.2-41.1,11.5c12.7,8.1,27.4,12.5,42.5,12.5c51,0,78.9-42.2,78.9-78.9c0-1.2,0-2.4-0.1-3.6 C182.7,97.4,189.2,93.7,192.9,88.1z"></path>
|
||
</g>
|
||
</g>
|
||
<circle class="st30" cx="124.4" cy="128.8" r="108.2"></circle>
|
||
</g>
|
||
</svg></a></li>
|
||
<li class="list-inline-item"><a href="https://www.facebook.com/nationallibraryofmedicine" aria-label="Facebook" rel="noopener noreferrer" target="_blank">
|
||
<svg xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink" version="1.1" x="0px" y="0px" viewBox="0 0 249 249" style="enable-background:new 0 0 249 249;" xml:space="preserve">
|
||
<style type="text/css">
|
||
.st10 {
|
||
fill: #FFFFFF;
|
||
}
|
||
|
||
.st110 {
|
||
fill: none;
|
||
stroke: #FFFFFF;
|
||
stroke-width: 8;
|
||
stroke-miterlimit: 10;
|
||
}
|
||
</style>
|
||
<title>Facebook</title>
|
||
<g>
|
||
<g>
|
||
<path class="st10" d="M159,99.1h-24V88.4c0-5,3.3-6.2,5.7-6.2h16.8V60l-24.4-0.1c-22.1,0-26.2,16.5-26.2,27.1v12.1H90v22.5h16.9 v67.5H135v-67.5h21.7L159,99.1z"></path>
|
||
</g>
|
||
</g>
|
||
<circle class="st110" cx="123.6" cy="123.2" r="108.2"></circle>
|
||
</svg>
|
||
</a></li>
|
||
<li class="list-inline-item"><a href="https://www.youtube.com/user/NLMNIH" aria-label="Youtube" target="_blank" rel="noopener noreferrer"><svg xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink" version="1.1" x="0px" y="0px" viewBox="0 0 249 249" style="enable-background:new 0 0 249 249;" xml:space="preserve">
|
||
<title>Youtube</title>
|
||
<style type="text/css">
|
||
.st4 {
|
||
fill: none;
|
||
stroke: #FFFFFF;
|
||
stroke-width: 8;
|
||
stroke-miterlimit: 10;
|
||
}
|
||
|
||
.st5 {
|
||
fill: #FFFFFF;
|
||
}
|
||
</style>
|
||
<circle class="st4" cx="124.2" cy="123.4" r="108.2"></circle>
|
||
<g transform="translate(0,-952.36218)">
|
||
<path class="st5" d="M88.4,1037.4c-10.4,0-18.7,8.3-18.7,18.7v40.1c0,10.4,8.3,18.7,18.7,18.7h72.1c10.4,0,18.7-8.3,18.7-18.7 v-40.1c0-10.4-8.3-18.7-18.7-18.7H88.4z M115.2,1058.8l29.4,17.4l-29.4,17.4V1058.8z"></path>
|
||
</g>
|
||
</svg></a></li>
|
||
</ul>
|
||
</div>
|
||
<div class="col-lg-3 col-12">
|
||
<p class="address_footer text-white">National Library of Medicine<br />
|
||
<a href="https://www.google.com/maps/place/8600+Rockville+Pike,+Bethesda,+MD+20894/@38.9959508,-77.101021,17z/data=!3m1!4b1!4m5!3m4!1s0x89b7c95e25765ddb:0x19156f88b27635b8!8m2!3d38.9959508!4d-77.0988323" class="text-white" target="_blank" rel="noopener noreferrer">8600 Rockville Pike<br />
|
||
Bethesda, MD 20894</a></p>
|
||
</div>
|
||
<div class="col-lg-3 col-12 centered-lg">
|
||
<p><a href="https://www.nlm.nih.gov/web_policies.html" class="text-white">Web Policies</a><br />
|
||
<a href="https://www.nih.gov/institutes-nih/nih-office-director/office-communications-public-liaison/freedom-information-act-office" class="text-white">FOIA</a><br />
|
||
<a href="https://www.hhs.gov/vulnerability-disclosure-policy/index.html" class="text-white" id="vdp">HHS Vulnerability Disclosure</a></p>
|
||
</div>
|
||
<div class="col-lg-3 col-12 centered-lg">
|
||
<p><a class="supportLink text-white" href="https://support.nlm.nih.gov/">Help</a><br />
|
||
<a href="https://www.nlm.nih.gov/accessibility.html" class="text-white">Accessibility</a><br />
|
||
<a href="https://www.nlm.nih.gov/careers/careers.html" class="text-white">Careers</a></p>
|
||
</div>
|
||
</div>
|
||
<div class="row">
|
||
<div class="col-lg-12 centered-lg">
|
||
<nav class="bottom-links">
|
||
<ul class="mt-3">
|
||
<li>
|
||
<a class="text-white" href="//www.nlm.nih.gov/">NLM</a>
|
||
</li>
|
||
<li>
|
||
<a class="text-white" href="https://www.nih.gov/">NIH</a>
|
||
</li>
|
||
<li>
|
||
<a class="text-white" href="https://www.hhs.gov/">HHS</a>
|
||
</li>
|
||
<li>
|
||
<a class="text-white" href="https://www.usa.gov/">USA.gov</a>
|
||
</li>
|
||
</ul>
|
||
</nav>
|
||
</div>
|
||
</div>
|
||
</div>
|
||
</section>
|
||
<script type="text/javascript" src="/portal/portal3rc.fcgi/rlib/js/InstrumentOmnitureBaseJS/InstrumentNCBIConfigJS/InstrumentNCBIBaseJS/InstrumentPageStarterJS.js?v=1"> </script>
|
||
<script type="text/javascript" src="/portal/portal3rc.fcgi/static/js/hfjs2.js"> </script>
|
||
</div>
|
||
</div>
|
||
</div>
|
||
<!--/.page-->
|
||
</div>
|
||
<!--/.wrap-->
|
||
</div><!-- /.twelve_col -->
|
||
</div>
|
||
<!-- /.grid -->
|
||
|
||
<span class="PAFAppResources"></span>
|
||
|
||
<!-- BESelector tab -->
|
||
|
||
|
||
|
||
<noscript><img alt="statistics" src="/stat?jsdisabled=true&ncbi_db=books&ncbi_pdid=book-part&ncbi_acc=NBK604916&ncbi_domain=nciprotocols&ncbi_report=classic&ncbi_type=fulltext&ncbi_objectid=&ncbi_pcid=/NBK604916/?report=classic&ncbi_pagename=Measurement of Nanoparticle Effects on Cytotoxic Activity of NK Cells by Label-Free RT-CES System - National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols - NCBI Bookshelf&ncbi_bookparttype=chapter&ncbi_app=bookshelf" /></noscript>
|
||
|
||
|
||
<!-- usually for JS scripts at page bottom -->
|
||
<!--<component id="PageFixtures" label="styles"></component>-->
|
||
|
||
|
||
<!-- CE8B5AF87C7FFCB1_0191SID /projects/books/PBooks@9.11 portal107 v4.1.r689238 Tue, Oct 22 2024 16:10:51 -->
|
||
<span id="portal-csrf-token" style="display:none" data-token="CE8B5AF87C7FFCB1_0191SID"></span>
|
||
|
||
<script type="text/javascript" src="//static.pubmed.gov/portal/portal3rc.fcgi/4216699/js/3879255/4121861/3501987/4008961/3893018/3821238/4062932/4209313/4212053/4076480/3921943/3400083/3426610.js" snapshot="books"></script></body>
|
||
</html> |