利用旋转式生物反应器快速扩增骨髓间充质干细胞  

Rapid expansion of bone marrow derived mesenchymal stem cells(MSCs) using rotary bioreactor

作　　者：倪明[1] 谢慧琪[2] 陈俊伟[1] 杨志明[2] 秦岭[1] 李刚[1,2] 

机构地区：[1]香港中文大学威尔斯亲王医院矫形与创伤医学系 [2]四川大学生物治疗国家重点实验室干细胞与组织工程研究室,成都610041

出　　处：《医用生物力学》2009年第1期28-33,共6页Journal of Medical Biomechanics

基　　金：国家自然科学基金资助项目(30872635/C160705)

摘　　要：目的探讨旋转式生物反应器是否能促进骨髓来源的间充质干细胞(MSCs)的快速扩增。方法将MSCs细胞放入盛有MyelocultTM培养液生物反应器中动态培养,并在MyelocultTM培养液中补充一些因子,如干细胞因子(SCF),白介素3和6(IL-3,IL-6)。检测和比较经反应器处理前后的MSCs的表型,增生和分化能力的变化。结果在生物反应器中培养8d,总细胞数、Stro-1+CD44+CD34-间充质干细胞和CD34+CD44+Stro-1-造血干细胞分别增加了9、29和8倍。实验研究结果显示,生物反应器扩增的间充质干细胞表达了原始间充质干细胞的标记物内皮因子(SH2)和波形蛋白,却没有发现与细胞系的分化有关的标记物,包括骨钙素(成骨指标),II型胶原(成软骨指标)和C/EBPα(成脂指标)。生物反应器组的细胞集落生成效率(成纤维细胞集落/天)是对照组(无生物反应器组)的1.44倍。经诱导后,生物反应器扩增的间充质干细胞可以分化成成骨细胞,软骨细胞和脂肪细胞,其分化能力与未经生物反应器处理的MSCs无区别。结论旋转式生物反应器结合改良的MyelocultTM培养液可用于快速扩增间充质干细胞。Objecthe To examine the feasibility of expanding MSCs from the isolated bone marrow mononuclear cells using a rotary bioreactor system. Methods The cells were cultured in a rotary bioreactor with Myelocult？ medium with addtion of supplementary factors such as stem cell factor （SCF）, interleukin 3 and 6 （IL-3, IL-6）. The MSCs were measured and compared the changed of its apparent, hyperplasia and differentiation before and after treated by the bioreactor. Resules After 8 days of culture, total cell numbers, Stro-1 ＋ CD44 ＋ CD34 MSCs and CD34 ＋ CD44＋ Stro-1- HSCs were increased 9,29, and 8 folds respectively. The bioreactorexpanded MSCs showed expression of primitive MSCs markers endoglin （SH2） and vimentin, whereas markers associated with lineage differentiation including osteocalcin （osteogenesis）, Type Ⅱ collagen （chondrogenesis） and C/EBPα （adipogenesis） were not detected. Colony forming efficiency-flbroblast per day （CFE-F/day） of the bioreactor-treated cells was 1.44-fold higher than that of the cells without bioreactor treatment. Upon induction, the bioreactor-expanded MSCs were able to differentiate into osteoblasts, chondrocytes and adipocytes, which showed no difference with that of MSCs without treating by Bioreactor. Conclusion It could conclude that the rotary bioreactor with the modified Myelocult？ medium reported in this study could be used to rapidly expand MSCs.

关 键 词：间充质干细胞 造血干细胞 生物反应器 分化 

分 类 号：R318.01[医药卫生—生物医学工程]