无血清培养基SYL-SF对间充质干细胞的培养和扩增作用  

Culture and expansion of mesenchymal stem cells by serum free medium

作　　者：张文成[1] 王韫芳[1] 常铭洋[1] 池木根[2] 孔维霞[3] 裴雪涛[1] 吴祖泽[2] 

机构地区：[1]军事医学科学院全军干细胞与再生医学重点实验室,北京100850 [2]军事医学科学院放射与辐射医学研究所,北京100850 [3]军事医学科学院附属307医院移植科,北京100071

出　　处：《中国医药生物技术》2012年第5期321-326,共6页Chinese Medicinal Biotechnology

摘　　要：目的 探讨新型无血清培养基SYL—SF对脐带间充质干细胞的体外培养和扩增作用。方法机械分离方法获得脐带间充质干细胞，从P1代开始将含小牛血清的原代培养基SYL-NBCS更换为实验组SYL-SF和对照组MSCM-SF两种无血清培养基对细胞进行培养、传代、扩增和维持。利用Alexa Fluor 488 annexin V检测细胞凋亡率，CCK-8法比较传代细胞的增殖效率，并对细胞进行形态、表型、染色体核型和分化能力等方面的检测和评价。结果分离得到的单个脐带问充质干细胞在原代培养基SYL-NBCS中培养8-15d可获得原代（P0）细胞，更换为无血清培养基后，SYL-SF组细胞凋亡率明显低于MSCM-SF组；在细胞传代过程中SYL—SF组细胞增殖能力较强且维持MSCs特异性表面标志和向脂肪、骨、软骨等多向分化的潜能，并且在传代过程中维持正常染色体核型。结论SYL—SF无血清培养基可以对UC-MSCs进行良好地体外扩增和维持。Objective To investigate the effect of a new serum-free medium SYL-SF on the culture and expansion of umbilical cord derived mesenchymal stem ceils （UC-MSCs）. Methods UC-MSCs were obtained from aborted human umbilical cord by mechanical isolation method, and were maintained in primary culture medium SYL-NBCS. From passage 1 （P1）, UC-MSCs were maintained and expanded in two different types of serum-free medium SYL-SF and MSCM-SF. Apoptosis was analyzed with Alexa Fluor 488 annexin V apotosis kit and cellular proliferation was analyzed with CCK-8 method. Cellular morphology, karyotype, cell surface marker and differentiation capacity were also detected. Results Primary UC-MSCs （P0） are obtained 8 to 15 days after the mechanical isolation and cultured in primary medium SYL-NBCS. When the cells are changed into serum-free medium, the apoptosis rate of the cells cultured in SYL-SF is significantly lower than those in MSCM-SF. In SYL-SF medium, UC-MSCs expend faster and maintain MSCs-specific surface markers as well as the multipotent differentiation potential towards fat, bone and cartilage. They also maintain normal karyotype after 8 passages of culture in SYL-SF. Conclusion SYL-SF as a novel serum-free medium can be used to support the expansion and maintenance of UC-MSCs in vitro.

关 键 词：间质干细胞 培养基 无血清 生物疗法 蛋白质类 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]