体外连续传代培养脐带间充质干细胞染色体核型的稳定性  

Karyotype stability of human umbilical cord mesenchymal stem cells during culture in vitro

作　　者：王军[1,2] 徐曼[1,2] 盛宏霞[1,2] 孔维霞[1,2] 扈江伟[1,2] 廖丽[1,2] 汤永永[1,2] 宁红梅[1,2] 张斌[1,2] 陈虎[1,2] 

机构地区：[1]军事医学科学院附属医院造血干细胞移植科 [2]军事医学科学院细胞与基因治疗中心,北京10007

出　　处：《军事医学》2012年第8期599-602,共4页Military Medical Sciences

基　　金：国家"863"计划资助项目(2011AA020114);国家自然科学基金资助项目(81070435)

摘　　要：目的分析不同代次人脐带间充质干细胞(umbilical cord mesenchymal stem cells,UC-MSCs)的染色体核型,初步评价UC-MSCs在体外连续传代培养过程中染色体结构的稳定性。方法采用胶原酶消化法分离UC-MSCs,贴壁培养传代,通过细胞形态、免疫表型及多向分化潜能等生物学特性进行鉴定,利用G显带分析第3、5、7代细胞的染色体核型。结果染色体核型分析显示,第3、5、7代UC-MSCs为正常二倍体核型,G显带未见染色体结构异常。UC-MSCs呈成纤维样形态生长,高表达CD73、CD90、CD105,不表达CD34、CD45、CD40、CD80、CD86、CD154、HLA-DR;在特定的体外诱导条件下可以向骨、脂肪、软骨分化。结论 UC-MSCs在体外连续传代培养7代以内染色体结构稳定,为临床应用UC-MSCs的安全性提供了遗传学方面的实验依据。Objective To analyze the karyotype of human umbilical cord mesenchymal stem cells （UC-MSCs） and to evaluate the stability of chromosome structure of UC-MSCs during continuous passaging in vitro. Methods The UC-MSCs were isolated by digestion with collagenase and subcuhured in vitro. They were characterized by morphology, immunophe- notype and multi-directional differentiation potential. Furthermore, the karyotype of passage 3 （ P3 ）, P5 and P7 UC-MSCs was analyzed by G-banding technique. Results The karyotype analysis of P3, P5 and P7 UC-MSCs showed a normal dip- loid karyotype with 46 chromosomes and there were no abnormal changes in chromosome structure. UC-MSCs exhibited fibroblastic morphology and were CD73, CD90, and CD105 positive and CD34, CD45, CD40, CDS0, CD86, CD154, and HLA-DR negative. Under special induction conditions, UC-MSCs could differentiate into osteocytes, adipocytes and chondro- cytes, Conclusion When cultured for 7 passages in vitro, UC-MSCs can maintain stable chromosome structure, which pro- vides an experimental basis for the safety of UC-MSCs eytotherapy.

关 键 词：间充质干细胞 脐带 核型 细胞增殖 连续传代 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]