脐血干细胞的分离方法比较

目的:为合理选择脐血干细胞的分离方法提供参考.方法:采用改良HES(羟乙基淀粉)沉降分离法和Ficoll分离法分离脐血干细胞,比较两种方法分离前后的有核细胞数、分离后的CD34 细胞数、分离后的有核细胞回收率及台盼蓝拒染率.结果:改良HES沉降分离法和Ficoll分离法分离前的脐血量、有核细胞浓度和细胞数差异无统计学意义(P＞0.05);分离后改良HES法有核细胞数、CD34 细胞数、有核细胞回收率高于Ficoll 分离法(P＜0.05),而显示有核细胞活力的台盼蓝拒染率两种方法比较差异无统计学意义 (P＞0.05).结论:改良HES分离法的有核细胞回收率高,分离速度快、污染机会小、终体积小,是脐血干细胞分离较好的方法.

Methods of Separating Stem Cells from Umbilical Cord Blood

Objective:To choose an appropriate method of separating stem cells from umbilical cord blood.Methods:Umbilical cord blood(UCB)samples were collected by blood bags and were separated by using hydroxyethyl starch(HES)and Ficoll method.Nuclear cells,CD34+ cells and trypan blue dye viability were compared between HES and Ficoll method.Results:There was no difference in the volume of umbilical cord blood,the density and the number of the nuclear cells between HES and Ficoll method before separating(P>0.05).But after separating,the number of nuclear cells and CD34+ cells,the rate of recollection for the nuclear cells(the percentage of the number of nuclear cells of post-separation and pre-separation)by using HES separating were higher than that of using Ficoll separating(P<0.05).There was no difference in the rate of refusing trypan blue dye between HES and Ficoll separating(P>0.05).Conclusions:The quantity of human cord blood-derived stem cells collected by using improved HES separation is high,the operation is simple and convenient,the speed of separating isquick,the opportunity of pollution is small and final volume is small,which is a better method of separating stem cells from umbilical cord blood.