人脐血CD34^+ CD38^-细胞免疫表型和染色体核型特征分析  

Karyotyping and immunophenotyping analyses of the CD34^+CD38^- cells isolated from human umbilical cord blood

作　　者：田虹[1] 郑金娥[2] 龚非力[1] 王兴兵[2] 黄士昂[2] 陈忠[3] 

机构地区：[1]华中科技大学同济医学院医学遗传室,武汉430030 [2]华中科技大学同济医学院附属协和医院干细胞中心 [3]美国犹他大学医学院儿科分子细胞遗传室

出　　处：《中华血液学杂志》2005年第5期257-260,共4页Chinese Journal of Hematology

基　　金：国家杰出青年基金资助项目(30225038);国家973基金资助项目(001CB510103)

摘　　要：目的分离脐血干/祖细胞(CD34+CD38-)进行体外长期培养,观察分析其增殖、细胞表面分子标志和染色体核型的特征。方法用流式细胞仪分选CD34FITC和CD38PE标记的CD34+CD38-脐血原始细胞,在含细胞生长因子IL3、IL6、GMCSF、EPO、SCF和胰岛素样生长因子的干细胞培养基中培养6个月,用流式细胞术检测体外培养30d的干/祖细胞表面标记,并用G显带方法分析其染色体核型。结果在一定培养条件下,经7～12d培养,脐血干/祖细胞(CD34+CD38-)开始增殖。培养6个月后,每孔接种1个细胞,细胞数增殖至250～350个;每孔接种10个细胞,细胞数可增殖至400～500个。每孔接种1个细胞其细胞增殖峰持续时间(8～9代)比接种10个细胞(6～7代)长。经体外长期培养增殖,细胞仍强烈显示干/祖细胞表面分子标记(CD34+CD38-);细胞染色体数目、结构未见异常。结论脐血干/祖细胞(CD34+CD38-)经体外特异性培养增殖,可为大量脐血干/祖细胞移植提供细胞来源。Objective To cultivate hematopoietic stem/progenitor cells (CD34 +CD38 -) isolated from umbilical cord blood (UCB) long termly for the observation of cell growth and expansion in vitro, surface marker expression, and chromosomal complements. Methods By flow cytometry CD34-FITC and CD38-PE labeled CD34 + and CD38 - stem/progenitor cells were isolated from UCB. The cells were cultivated in vitro for 6 months in a stem cell culture system with addition of six kinds of cell growth factors (IL-3, IL-6, GM-CSF, Epo, SCF, IGF-1). One month after cultivation, cultured cells were investigated for surface marker expression by flow cytometry and karyotype by G banding method. Results After 7~12 days cultivation, the CD34 +CD38 - stem/progenitor cells began proliferation. The proliferation rate and the peak proliferation duration were greater in 1 cell/well cultivation conditions than in 10 cells/well. The cells remained CD34 +CD38- and their karyotypic characteristics remained unchanged. Conclusion CD34 +CD38 - stem/progenitor cells from UCB may provide a larger than original amount of stem/progentor cells for transplantation after long-term cultivation in vitro.

关 键 词：人脐血 CD34^+CD38^-细胞免疫表型 染色体核型 造血干细胞 

分 类 号：R394[医药卫生—医学遗传学]