用条件培养液代替重组因子培养小鼠高增殖潜能集落形成细胞

利用富含ＩＬ－３的ＷＥＨＩ－３细胞条件培养液（ＷＥＨＩ３－ＣＭ）和富含Ｍ－ＣＳＦ的Ｌ９２９细胞条件培养液（Ｌ９２９－ＣＭ）代替重组因子在体外用单层琼脂法培养小鼠ＨＰＰ－ＣＦＣ获得成功。在合用ＷＥＨＩ３－ＣＭ和Ｌ９２９－ＣＭ的基础上，联合ｒｈＩＬ－１，ｒｈＩＬ－６和ｒｍＧＭ－ＣＳＦ，ＨＰＰ－ＣＦＣ的产率有所提高。用５－ＦＵ处理小鼠２天后，骨髓细胞中ＨＰＰ－ＣＦＣ得以浓集。产生的ＨＰＰ－ＣＦＣ集落直径

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.