脐血单个核细胞体外红系定向扩增和分化的研究

Studies on ex vivo production of erythroid cells derived from humancord blood MNC expansion and committed differentiation

目的 :探讨利用脐血单个核细胞 (MNCs)体外定向诱导扩增和分化为成熟红细胞以期作为临床输血源的可行性。方法 :10例脐血MNC在无血清无基质的液体培养条件下 ,在含SCF、IL 3、EPO和地塞米松 (DEX)等因子不同组合的A(空白对照 )、B(SCF加EPO)、C(SCF加EPO加IL 3)和D(SCF加EPO加IL 3加DEX)四组进行培养 14d。结果 :在第 10天时 ,B组、C组和D组中红系祖细胞扩增数均达到最高点 ,而含有SCF加IL 3加EPO加DEX的D组中红系祖细胞扩增了 4 .6± 0 .4倍 ,明显高于其他三组 (P <0 .0 1) ;结论 :①脐血MNC在含有SCF加IL 3加EPO加DEX的因子组合的无血清培养中能使红系祖细胞得到有效的扩增 ,且红系祖细胞扩增数在第 10天时达到最高。②采用分步的培养方法不仅获得更多更纯的红系细胞 ,而且能加速红系的分化 ,获得更高比例的成熟红细胞。

Objective:To evaluated the feasibility of using the red blood cells produced fuom induction of committed proliferation and differentiation of cord blood MNC ex vivo as a new sources of transfusion.Method:10 samples of cord blood mononuclear cells were incubated for 14 days in serum-free liquid suspansion culture system(StemSpen) with various cytokine combinations as following:negtive control(groupA),SCF+EPO(group B),EPO+SCF+IL-3(group C) and SCF+EPO+IL-3+DEX(group D).Result:At 10 days , the number of CFU-Es increased to the highest level in group B,group C and group D, then there were (4.6)±(0.4) folds expansion for CFU-E in group D,which were the highest in four groups(P<(0.01)).Conclusion:Firstly,in the group of SCF+IL-3+EPO+DEX , erythroid progenitors could greatly be expanded from cord blood MNCs in the serum-free liquid culture system.Secondly, the method of two-step culture in the serum-free liquid culture system makes cord blood MNCs not only prolifterate greatly but also differentiate deeply to mature erythroid cells,which will be a new sources of clinical transfussion.