Human amnion mesenchymal stem cells (hAMSCs) are an attractive cell source for bone tissue engineering. However, their utility is restrained by low expansion rate and poor osteogenic differentiation under static culture conditions. In the present study, hAMSCs were seeded and cultured in 12-wells plates as static culture system or Cytodex3 microcarrier-based dynamic culture system. Both proliferation and osteogenic differentiation of hAMSCs in spinner flasks were investigated and compared with those in static plates. Total cell number, alkaline phosphatase activity, and calcium deposition were determined. The effects of fluid flow stimulation and collagen coating on osteogenic differentiation of hAMSCs were further analyzed in static plates. The results show that higher cell density and stronger osteogenic differentiation in microcarrier-based dynamic culture were achieved than those in static plates. It was further found that fluid flow stimulation and collagen coating could promote osteogenic differentiation of hAMSCs, and there exists additive effects when fluid flow stimulation and collagen coating were combined. Therefore, Cytodex 3 microcarrier-based dynamic culture system represents a promising strategy for expansion and osteogenic differentiation of hAMSCs in an integrated manner.%人羊膜间充质干细胞(hAMSCs)作为一种多潜能干细胞有望应用于骨组织工程.但是,传统的静态培养和诱导方法并不利于间充质干细胞的体外扩增和成骨分化.实验通过考察和比较在孔板中静态及微载体动态培养条件下hAMSCs的扩增与成骨诱导,以建立集间充质干细胞增殖和成骨分化诱导为一体的动态培养体系.将hAMSCs接种到Cytodex 3微载体上,计细胞数,检测成骨诱导分化后细胞的碱性磷酸酶活性与钙基质含量,与二维静态培养系统比较;另外,基于二维培养体系进一步考察了流体刺激和胶原基质修饰对hAMSCs成骨分化的影响.结果表明:在微载体动态诱导培养条件下可获得较高的细胞数和较好的成骨诱导分化效果,提高了碱性磷酸酶活性与钙基质含量;流体刺激与包被的Ⅰ型胶原均可促进hAMSCs的成骨分化,且两者联合应用时促进作用更明显.因此,基于微载体的动态培养体系可以促进间充质干细胞在骨组织再生工程中的应用.
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