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Published on: July 24, 2014
[Culture of murine HPP-CFC in vitro using conditioned medium substitutefor recombinant growth factors]
Objects:
To study the clonal growth on single layer, agar of high proliferative potential colony-forming cells (HPP-CFC) of bone marrow obtained from both normal mice (NBM) and mice treated 2 days earlier with 5-fluorouracil (FU,BM) using conditioned media.
Material And Methods:
Using liquid cell culture to harvert WEHI2-CM and L929-CM as the substitutes for IL-3 and M-CSF respectively; using semi-solid culture to assay HP-CFCin vitro.
Results:
(1) the combination of WEHI3-CM and L929-CM produced HPP-CFC in vitro, and the addition of rhIL-1 alpha, rhIL-6 and rmGM-CSF further enhanced HPP-CFC was enriched in the FU2 BM. Morphological investigation showed that the HPP-CFC colony was large and compact, great than 0.5mm in diameter and contained more than 50,000 cells, most of which were macrophage like cells. The hydroxyurea suicide rate for HPP-CFC was less than 10%.
Conclusion:
The results suggested that (1) WEHI3-CM and L929-CM can be used as the substitutes of IL-3 and M-CSF to stimulate the formation of HPP-CFC; (2) FU can enrich HPP-CFC of the mice.
