组织块法分离人脐带间充质干细胞的研究  

The research of hunman unmbilical cord derived-mesenchymal stem cell by tissue isolation

作　　者：李铎[1] 石钏 洪敬欣 孟恒星 韩俊领[1,2] 张宇光[1,2] 

机构地区：[1]中国医学科学院北京协和医学院血液学研究所,天津300020 [2]协和干细胞基因工程有限公司,天津300384

出　　处：《中国医药导报》2011年第24期19-22,共4页China Medical Herald

摘　　要：目的:应用组织块法分离人脐带间充质干细胞(hUC-MSCs)并对其生物学特性进行研究。同时探讨在不同储存条件下的脐带是否对组织块法的分离成功率产生影响。方法:无菌条件下采集足月儿的脐带,应用组织块法分离hUC-MSCs,通过流式细胞术观察其免疫学、生长特性。脐带在常温条件和冷冻条件下储存后进行组织块法分离,观察细胞生长情况。结果:应用组织块法能获得贴壁生长的hUC-MSCs,流式细胞仪分析结果显示细胞高表达常见MSC表面分子CD105、CD73及HLA-ABC;低表达造血细胞表面标记CD34、CD45及HLA-DR。常温储存条件下,随着处理时间的延长,获得原代细胞数量下降;组织冷冻后可以获得hUC-MSCs。结论:组织块分离法可以获得贴壁生长的hUC-MSCs,并成功传代扩增;脐带采集后24 h内处理,hUC-MSCs获得数量及分离成功率高;随着处理时间延长,hUC-MSCs获得数量减少,分离成功率降低。Objective：To isolate human umbilical cord mesenchymal stem cells（hUC-MSCs） by tissue isolation,and to explore the impact of isolating by tissue isolation in different storage condition.Methods：Human umbilical cord was processed in different storage condition with tissue isolation.The cellular growth and immunological characteristics of hUC-MSCs by flow cytometry was observed.Results：hUC-MSCs could be successful obtained by tissue isolation.Flow cytometry analysis showed that hUC-MSCs highly express cell surface molecules such as CD105,CD73,HLA-ABC,while low-grade expressed the cells surface markers like CD34,CD45 and HLA-DR.The primary cell number of hUC-MSCs decreased as the treatment time elongation,and we could obtained hUC-MSCs from frozen cord pieces.Conclusion：hUC-MSCs can be successfully obtained by tissue isolation,and there is different among different treatment time group,but no significant difference between cryopreservation group and non-cryopreservation group within 24 h.

关 键 词：人脐带间充质干细胞 组织块分离法 生物学鉴定 

分 类 号：R394.2[医药卫生—医学遗传学]