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CD34~+脐血造血干细胞的体外培养及其生物学特性研究
引用本文:胡卫华,王净,何向锋,褚莉莉,赵枫姝,蒋翠莲,李晓丽. CD34~+脐血造血干细胞的体外培养及其生物学特性研究[J]. 皖南医学院学报, 2010, 29(3): 161-165
作者姓名:胡卫华  王净  何向锋  褚莉莉  赵枫姝  蒋翠莲  李晓丽
作者单位:1. 东南大学医学院病原生物学和免疫学系,江苏南京210009;皖南医学院遗传医学研究室,安徽芜湖241002
2. 东南大学附属中大医院妇产科,江苏南京,210009
3. 东南大学医学院病原生物学和免疫学系,江苏南京,210009
4. 南京儿童医院儿科研究所,江苏南京,210009
基金项目:安徽省高校省级自然科学研究项目 
摘    要:目的:建立稳定的CD34+脐血造血干细胞(CD34+UHSC)的体外分离、培养扩增体系,并研究其生物学特性。方法:密度梯度离心及免疫磁珠法从脐血中分离CD34+UH-SC,加入细胞因子SCF、IL-3和IL-6进行体外培养扩增,观察细胞形态及其生长特性。应用流式细胞仪测定细胞周期及其CD34分子的表达,研究其增殖、生长和分化特性。以RT-PCR法检测干细胞高表达基因ABCG2的转录表达。结果:在体外培养条件下,CD34+UHSC悬浮生长,约1周后易形成克隆球,增殖旺盛,呈现指数生长期;细胞增殖周期随体外培养时间延长而逐渐趋缓;CD34+UHSC表面标志亦随体外培养时间延长而表达下降,同时ABCG2基因的表达亦逐步下调。结论:体外获得纯化的CD34+UHSC,生长稳定、增殖能力强,但随体外培养时间的延长,CD34+分子标志逐步丢失,提示在增殖旺盛的指数生长期7~21d内最适用于各项相关实验研究。

关 键 词:脐血  CD34~+造血干细胞  体外培养  流式细胞术

Culture of CD34+ hematopoietic stem cells derived from human umbilical cord blood and their biological properties in vitro
HU Wei-hua,WANG Jing,HE Xiang-feng,CHU Li-li,ZHAO Feng-shu,JIANG Cui-lian,LI Xiao-li. Culture of CD34+ hematopoietic stem cells derived from human umbilical cord blood and their biological properties in vitro[J]. Acta Academiae Medicinae Wannan, 2010, 29(3): 161-165
Authors:HU Wei-hua  WANG Jing  HE Xiang-feng  CHU Li-li  ZHAO Feng-shu  JIANG Cui-lian  LI Xiao-li
Affiliation:HU Wei-hua,WANG Jing,HE Xiang-feng,CHU Li-li,ZHAO Feng-shu,JIANG Cui-lian,LI Xiao-li Department of Pathogenic Biology and Immunology,Medical College,Southeast University,Nanjing 210009,China
Abstract:Objective:To establish a stable culture system in vitro for isolating CD34+ hematopoietic stem cells derived from human umbilical cord blood (CD34+UHSCs) and to investigate their biological characteristics.Methods:Density-gradient centrifugation and immunomagnetic beads separation methods were used to derive CD34+UHSCs from human umbilical cord blood,cultured with the cytokines of SCF,IL-3 and IL-6 and amplification in vitro to observe the cell morphology and biological properties.Flow cytometry was used to...
Keywords:umbilical cord blood  CD34 hematopoietic stem cells  culture in vitro  flow cytometry  
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