人羊膜间充质干细胞的分离_培养及鉴定.pdfVIP

第 卷第 期 遵 义 医 学 院 学 报 32 3 Vo1.32 No.3 年 月 2009 6 ACTA ACADEMIAE MEDICINAE ZUNYI Jun. 2009 人羊膜间充质干细胞的分离、培养及鉴定 方 宁，张 路，宋秀军，章 涛，陈代雄 （遵义医学院附属医院贵州省细胞工程重点实验室，贵州遵义563000） [摘 要] 目的 建立人羊膜间充质干细胞分离、培养及鉴定方法。方法 采用低速旋转-胰蛋白酶-胶原酶消化法 分离人羊膜间充质干细胞（hAMSCs），采用免疫细胞化学和流式细胞术分析其表型特征，使用含10%FBS 低糖- 7 DMEM 培养基培养。结果 从羊膜分离的hAMSCs 数为 (6.72±1.21)×10 /份（n=12），所分离的hAMSCs 明显表达间充 质细胞标志物波形蛋白，不表达上皮细胞标志物角蛋白CK19，高表达CD29 和CD44。培养 10 天hAMSCs 数量可增 殖167 倍。结论 建立了人羊膜间充质干细胞分离、培养及鉴定方法，hAMSCs 具有骨髓间充质干细胞的表型特征。 [关键词] 羊膜间充质细胞; 分离；培养；鉴定 [中图分类号] R394.2 [文献标识码] A [文章编号] 1000-2715 （2009）03-0234-03 Isolation, culture and identification of the mesenchymal stem cells derived from human amnion FANG Ning, ZHANG Lu, SONG Xiu-jun, ZHANG Tao, CHEN Dai-xiong (Key laboratory cell engineering of Guizhou Province, First Affiliated Hospital ,Zunyi Medical College, Guizhou Zunyi 563000, China) [Obstract] Objectives To investigate the isolation and culture condition of human amniotic mesenchymal stem cells (hAMSCs) and its phenotypic identification. Methods The hAMSCs were isolated from human amnion digested with trypsin -collaenase.The phenotype of hAMCs was identified by Immunocytochemistry and flow cytometry. The cells were cultured in LG-DMEM medium containing 10% fetal bovine serum. Results A term amnion may yield up to (6.72±1.21)× 7 10 hAMSCs by low velocity-trypsin-collaenase digestion . Vementin 19 was expressed in the AMSCs derived from human amnion, while was negative. CD29 and CD44 were highly expressed in the primary cultured hAMSCs. The total cell number of hAMSCs was expanded by 167 -fold after