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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 |
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| 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 |
| Institution: | 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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