人正常骨髓CD34~ 造血细胞体外扩增形成红系及混合系造血祖细胞的能力

根据阳性选择策略，采用ＣＩＭＳ－１００免疫磁性无菌分离系统富集人正常骨髓ＣＤ３４＋造血细胞，然后将其在含Ｅｐｏ＋ＧＭ－ＣＳＦ＋ＩＬ－３＋ＩＬ－６＋ＳＣＦ（简ＥＧＩＳ）组合造血生长因子的无基质液培体系中扩增４周，定期进行单个核细胞计数、ＢＦＵ－Ｅ及ＣＦＵ－Ｍｉｘ的培养。经ＦＡＣＳ鉴定所富集的ＣＤ３４＋造血细胞纯度平均＞９０％。人正常骨髓ＣＤ３４＋造血细胞在该条件下均可持续产生大量单个核细胞，最高可扩增１７７０倍。与ＨＰＰ－ＣＦＣ和ＣＦＵ－ＧＭ相反，ＢＦＵ－Ｅ和ＣＦＵ－Ｍｉｘ扩增量较低，最高仅为２．３６和２．４倍，且仅在第１周出现，随后迅速减少至停止。

EX VIVO EXPANSION OF BOTH BURST FORMING UNITS ERYTHROID AND COLONY FORMING UNITS MIXED FROM CD34 HEMATOPOIETIC CELLS OF HUMAN BONE MARROW

CIMS 100 immunomagnetic isolator, a new sterilized separation systems, was used to enrich CD34 hematopoietic cells from human bone marrow according to the positive selective strategy. Highly purified CD34 hematopoietic cells were cultured in stroml cells free liquid culture system(SCFLCS) supplemented with EPO GM-CSF IL 3 IL 6 SCF(EGIIS) every 48 hours. The cells harvested from SCFLCS were counted and assayed for MNC, BFU E and CFU Mix at weekly intervals for a period of 4 weeks. More than 90% purity of CD34 hematopoietic cells on average determined by flow cytometry was obtained. Under SCFLCS conditions, with the additions of EGIIS, CD34 hematopoietic cells from all 5 cases were able to produce continuously a significant amouts of MNC with the highest peak of 1770 folds during the periods of 4 weeks compared to input cells at day 0. In contrast, cells harvested from SCFLCS resulted in a slight expansion of both BFU E(2 36 fold) and CFU Mix(2 4 fold) compared to those of CFU GM and HPP CFU. The expansions of BFU E and CFU Mix only occurred during the first week of cultivation, then there was a rapid decrease or disappearance of both BFU E and CFU Mix.