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

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

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。

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.