脐血CD34+细胞的分离与纯化

Separating & Purifying CD34+ Cell from Umbilical Cord Blood

脐血造血干细胞的分离是进行干细胞移植、体外扩增培养的关键。研究采用明胶自然沉降法、Ficoll分层法分离脐血MNC后,用MACS分离纯化CD34+细胞,计数并用流式细胞仪进行纯度分析。结果显示:每份脐血的采集量平均为95+52ml,3g/dl明胶自然沉降法所获MNC密度平均为(5.76±0.67)×107/ml;CD34+细胞密度平均为(5.53±1.16)×105/ml,较Fi-coll分层法高(P<0.01)。因此,3g/dl明胶自然沉降法是一种较理想的分离MNC的方法,用明胶沉降法和MACS相结合是分离脐血CD34+细胞的理想方法。

Objective Separating & purifying CD34+ cell from umibilical cord blood is a hinge of stem celi transfer and in-vitro culture MNC were isolated from umbilical cord blood by using denshy grodlent centrifugation(Ficoll-Hypaque)or 3 g/dl gelatin matural sedimentation methods,then separating CD34+ cell from MNC by MACS.Results show,the number of MNC and CD34+ cell were(5.76±0.67)×107/ml and(5.53±1.16)×106/ml respectively by 3g/dl gelstin natural sedimentation.which was better than that from Ficoll-Hypaque method.So,3g/dl gelatin natural sedimentation is an optimal method for separating MNC.collaboration with MACs can separate and purify CD34+ surccessfully.