脐带血血清培养体系扩增脐带间充质干细胞及其生物学特征的实验研究  

Study on the Expansion of Umbilical Cord MSCs in Umbilical Cord Serum Cultural System and its Biological Characteristics

作　　者：葛殿华[1] 马红梅[2] 周学武[1] 呼晓[2] 冯丽艳[1] 雷娟[1] 杨淑梅[1] 毛慧玲[1] 宁彬[1] 

机构地区：[1]大庆龙南医院 [2]哈尔滨医科大学大庆校区,黑龙江大庆163319

出　　处：《黑龙江医药》2009年第6期799-801,共3页Heilongjiang Medicine journal

摘　　要：目的:通过观察以人脐带血血清为主体的培养体系对脐带间充质干细胞(Mesnchymal Stem Cells,MSCs)体外培养扩增的影响,探讨人脐带血血清替代动物血清用于培养临床组织工程用干细胞的可行性。建立从足月脐带分离间充质干细胞和体外传代培养技术,并对其生物学特性进行研究。方法:采用双酶法分离脐带间充质干细胞,并通过传代进行纯化和扩增培养,绘制生长曲线:用流式细胞仪检测脐带表面抗原及细胞周期。结果:成功建立了以人脐带血血清为主体的培养体系的脐带间充质干细胞培养扩增的方法:流式细胞仪检测结果显示,贴壁细胞均表达CD44、CD29,低表达CD106,不表达造血细胞表型CD14、CD34、CD45和内皮细胞表型CD31,也不表达HLADR:细胞倍增时间为30h,细胞周期分析表明,G0～G1期和+G2+M期所占比例分别为78.84%和11.16%。结论:应用灭活脐带血清培养体系可成功扩增人脐带间充质干细胞,培养的细胞具有间充质干细胞的基本特性,为建立间充质干细胞库和临床应用提供了理论依据。Objective:To observe the effect of ex-vivo expansion of umbilical cord MSCs after cultivated by umbilical cord serum cultural system,to explore the feasibility of umbilical cord serum substitute for animal blood serum be used to cultivate clinical organization engineering stem cells.Established the technique of separation MSCs from full-term umbilical cord and serial subcultivation in vitro.Studied their biological characteristics.Methods:Isolated umbilical cord MSCs with Dualenzyme method,purified and expanded through serial subcultivation in vitro.Drew the growth curve:Flow cytometry was used to detect the umbilical cord surface antigens and cell cycle.Results:Successfully established a expansion method of umbilical cord MSCs cultivated by mainly containing umbilical cord serum culture system.the flow cytometry showed that all adherent layer expressed CD44,CD29,lowly expressed CD106,did not express the phenotype CD14,CD34,CD45 of hematopoietic cells and endothelial cells phenotype CD31,,Also did not express HLA-DR;cell doubling time was 30h,cell cycle analysis showed,the percentage of G0～G1 phase and+G2+M phase was 78.84%and 11.16%.respectively.Conclusion:Inactivation cord serum culture system can be successfully expand human umbilical cord MSCs,it has the basic characteristics of MSCs,it offer the theory basis for establish MSCs bank and clinical application.

关 键 词：脐带血血清 间充质干细胞 生物学特征 

分 类 号：R457.2[医药卫生—治疗学]