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Review

Mouse Granulocyte-Macrophage Colony-Forming Unit Assay: Version 4

Barry W. Neun  1 , Edward Cedrone  1 , Marina A. Dobrovolskaia  1
In: National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols [Internet]. Bethesda (MD): National Cancer Institute (US); 2005 May. NCL Method ITA-3.
2020 May.
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Review

Mouse Granulocyte-Macrophage Colony-Forming Unit Assay: Version 4

Barry W. Neun et al.
Free Books & Documents

Excerpt

Myelosuppression is a common dose limiting toxicity of cytotoxic oncology drugs. Nanoparticles may distribute to bone marrow and/or release drug that is delivered to bone marrow. Therefore, understanding potential toxicity of nanoparticles or drugs which nanoparticles carry is an important step in preclinical safety evaluation.

Hematopoietic stem cells of bone marrow (BM) proliferate and differentiate to form discrete cell clusters or colonies. This document describes a protocol for quantitative analysis of granulocyte-macrophage (GM) colony-forming units (CFU), employing murine BM. This protocol can be used for both in vitro and ex vivo analyses. The in vitro protocol involves isolation of bone marrow cells from healthy animals, followed by treatment in vitro with nanoparticle formulations. In the ex vivo version, the bone marrow is isolated from animals injected with the nanoparticle formulation. The in vitro protocol does not account for nanoparticle biodistribution; however, in cases when dose information is not available and nanoparticle formulation is in the early phase of development, the in vitro protocol allows for rapid screening of potentially toxic nanoparticle formulations. The in vitro protocol can also give a quick estimation of the myelosuppressive potential of a cytotoxic oncology drug bound to a nanoparticle surface in comparison to a traditional formulation of the same cytotoxic oncology drug (for an example, see Figure 1).

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References

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