微载体悬浮培养成人骨髓间充质干细胞  

Cultivation of Human Mesenchymal Stem Cells on Macroporous CultiSpher G Microcarriers

作　　者：吴清法[1] 吴祖泽[1] 董波[1] 王立生[1] 

机构地区：[1]北京放射医学研究所

出　　处：《中国实验血液学杂志》2003年第1期15-21,共7页Journal of Experimental Hematology

基　　金：国家重点基础研究发展规划项目编号G19990 5 3 9;国家高技术研究发展计划资助项目编号2 0 0 1AA2 16161

摘　　要：本研究采用微载体旋转培养系统和常规静止培养系统对成人骨髓间充质干细胞 (mesenchymalstemcell,MSC)的培养进行比较。MSC是贴壁依赖性细胞 ,旋转培养系统采用CultiSpherG大孔微载体 ,浓度为 1g L ,常规静止培养在 12孔培养板中进行。两系统细胞接种密度均为 5× 10 4 cells ml。结果 :旋转培养 7天后达到最大活细胞密度5 .15 0× 10 5cells ml,常规静止培养第 5天就达到最大活细胞密度 1.6 75× 10 5cells ml。在微载体旋转培养中生成乳酸12 .0 6mmol L ,而常规静止培养中生成乳酸 13.10mmol L ,葡萄糖消耗分别为 7.38mmol L和 5 .37mmol L。在微载体旋转培养中平均乳酸产率为 1.6 3,远低于常规静止培养的平均乳酸产率 2 .4 4。这些表明 ,在微载体悬浮培养条件下 ,MSC生长更为旺盛 ,细胞产量更高 ,葡萄糖消耗和能量利用率优于常规静止培养。微载体悬浮培养 12天后 ,MSC依然保持其干细胞特性。结论Adult human bone marrow derived mesenchymal stem cells (MSCs) were cultured on microcarriers in spinner flasks and were compared with those in conventional culture in 12 well plates. For the production of adherently growing MSCs, macroporous CultiSpher G gelatin microcarriers were used in concentration of 1 g/L. The cells were seeded in a density of 5×10 4 cells/ml in both spinner culture and conventional stationary culture. The result showed that after 7 days of cultivation a maximum viable cell concentration of 5.15×10 5 cells/ml was obtained in spinner culture. Whereas the cell density increased to a maximum of 1.675×10 5 cells/ml on day 5 in conventional stationary culture . Lactate was produced up to 12.06 mmol/L in spinner culture and up to 13.10 mmol/L in stationary culture, and glucose was consumed up to 7.38 mmol/L and 5.37 mmol/L respectively. The average lactate yield on glucose consumption in spinner culture was only 1.63, lower than that in stationary culture 2.44. This indicated that the energy metabolism in spinner culture was significantly more efficient than that in conventional culture. After spinner culture for 12 days, the MSCs maintain the characteristics of stem cells. It is concluded that the microcarrier culture system is a suitable way to expand the seeding cells for tissue engineering.

关 键 词：间充质干细胞 细胞培养 微载体 组织工程 

分 类 号：R329－33[医药卫生—人体解剖和组织胚胎学]