Wu Qing-Fa, Wu Chu-Tse, Dong Bo, Wang Li-Sheng
Beijing Institute of Radiation Medicine, Beijing 100850, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2003 Feb;11(1):15-21.
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 x 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 x 10(5) cells/ml was obtained in spinner culture. Whereas the cell density increased to a maximum of 1.675 x 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.
将成人骨髓间充质干细胞(MSCs)接种于转瓶中的微载体上进行培养,并与传统的12孔板培养进行比较。为了培养贴壁生长的MSCs,使用浓度为1 g/L的大孔CultiSpher G明胶微载体。在转瓶培养和传统的静止培养中,细胞接种密度均为5×10⁴个细胞/ml。结果显示,培养7天后,转瓶培养获得的最大活细胞浓度为5.15×10⁵个细胞/ml。而在传统静止培养中,细胞密度在第5天增加到最大值1.675×10⁵个细胞/ml。转瓶培养中乳酸产量高达12.06 mmol/L,静止培养中高达13.10 mmol/L,葡萄糖消耗分别高达7.38 mmol/L和5.37 mmol/L。转瓶培养中基于葡萄糖消耗的平均乳酸产量仅为1.63,低于静止培养中的2.44。这表明转瓶培养中的能量代谢比传统培养显著更高效。转瓶培养12天后,MSCs保持干细胞特性。结论是微载体培养系统是一种适合用于组织工程中扩增种子细胞的方法。