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.
出处
《中国医药生物技术》
CSCD
2012年第5期321-326,共6页
Chinese Medicinal Biotechnology
关键词
间质干细胞
培养基
无血清
生物疗法
蛋白质类
Mesenchymal stem cells
Culture media, serum-free
Biological therapy
Proteins
