Colony formation in agar: in vitro assay for haemopoietic stem cells
article
Using a method in which embryo fibroblasts were used as feeder layers, the colony forming capacity in agar of a variety of mouse haemopoietic suspensions was compared with their CFUs content. A striking parallelism between the results of the two assays was found. In addition, under certain conditions higher numbers of CFUs could be retrieved from 5-day-old agar colonies than were originally plated, indicating that the CFCa (Colony Forming Cell agar) may fulfil the requirements of pluripotency as well as of self-renewal, both prerequisites for any haemopoietic stem cell candidate. Although our data by no means provide direct proof that the CFCs and the CFCa are identical, they certainly support such a concept. the contradictory findings by others that CFUs and CFUc (Colony Forming Unit culture) can be separated on a velocity gradient is attributed to different culture conditions, in other words, that their CFUcè are not identical with our CFUa.
Our findings also indicate that for mouse cells our soft agar colony assay meets the criteria of a quantitative assay for haemopoietic stem cells and that extension of this technique to bone marrow of primates including humans seems to be justified.
Our findings also indicate that for mouse cells our soft agar colony assay meets the criteria of a quantitative assay for haemopoietic stem cells and that extension of this technique to bone marrow of primates including humans seems to be justified.
TNO Identifier
353760
Source
Cell and Tissue Kinetics, 4(5), pp. 463-477.
Pages
463-477
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