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4.9. Colony-Forming Cell (CFC) Assay

ES Evgenia Sarrou
LR Laura Richmond
RC Ruaidhrí J. Carmody
BG Brenda Gibson
KK Karen Keeshan
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Methocult GF M3434 (Stem Cell Technologies, Vancouver, BC, Canada) methylcellulose media containing recombinant murine stem cell factor (rmSCF), recombinant murine interleukin-3 (rmIL-3), recombinant human interleukin-6 (rhIL-6) and recombinant human erythropoietin (rhEPO) was used. Equal numbers of BM c-kit-enriched, GFP-sorted cells were prepared in triplicate and resuspended in culture media. Cell suspension was added to methylcellulose and transferred to 3 × 35 mm dishes (Greiner-Bio-One, Gloucestershire, UK) for 10–20 days. Colonies were scored, and cells were analysed by flow cytometry. If required, cells were replated to a second round of CFC following the same protocol.

Copyright and License information:  ©2020 by the authors.
Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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