无血清培养体系原代培养脐带间充质干细胞

Primary culture of umbilical cord-derived mesenchymal stem cells in serum free medium

背景: 以含血清培养基培养的脐带间充质干细胞,存在着进一步应用的障碍。目的: 建立无血清培养体系原代培养脐带间充质干细胞。方法: 取脐带华通氏胶(Wharton’sJelly),采用机械法将组织胶切碎,1-3mm3大小,分别培养到含胎牛血清完全培养液中以及无血清培养液中。培养第11,14,17天收获细胞并进行相关检测。在符合2006年制定的干细胞最低评估标准的基础上,以集落形成单元指标评估何种培养体系能获得较多高质量的原代细胞。结果与结论: 无血清培养体系相对于经典的含血清的培养体系而言,细胞生长速度更快。另外,培养第11天,集落形成实验表明无血清培养体系下能获得更多的集落。因此,无血清培养体系可以保持间充质干细胞的特性,为替代含血清培养体系进行脐带间充质干细胞原代培养提供了可能。

OBJECTIVE： To establish serum free medium system for primary culture of umbilical cord-derived mesenchyma stem cells. METHODS： Wharton＇s Jelly was isolated from umbilical cord, minced, 1-3 mm^3, and subsequently incubated in either serum containing medium or serum free medium. Some cells were harvested on days 11, 14 and 17 for some detection. Based on the minimal criteria established by the International Society for Cellular Therapy in 2006, mesenchymal stem cells were assayed with colony forming unit-fibroblast. RESULTS AND CONCLUSION： The mesenchymal stem cells grew rapidly in serum free medium condition compared with serum containing medium. On day 11, the number of colonies was larger in serum free medium condition than that in serum containing medium. Thus, serum free medium could maintain properties of mesenchymal stem cells and provide possibility a credible alternative to serum containing medium for primary culture of umbilical cord-derived mesenchymal stem cells.