摘要
目的 筛选关节软骨组织工程种子细胞优化获取的高效促增殖剂。 方法 采用培养细胞3H -胸腺嘧啶脱氧核苷 (3H -TdR)掺入实验、四甲基偶氮唑蓝 (MTT)代谢检测及Ⅰ、Ⅱ型胶原mRNA逆转录聚合酶链反应 (RT -PCR)分析方法 ,比较一定浓度维生素C(vitaminC ,VC) +骨形态发生蛋白 2 (bonemorphogeneticprotein 2 ,BMP2 )、类胰岛素生长因子 1(insulin -likegrowthfac tor1,IGF1 ) (体积分数为 2 %的血清培养时 )及碱性成纤维细胞生长因子 (basicfibroblastgrowthfactor,bFGF)、转化生长因子 - β1 (transforminggrowthfactor- β1 ,TGF - β1 ) (体积分数为 10 %的血清培养时 )对适宜密度接种、体外单层传代培养兔关节软骨细胞的促增殖作用。 结果 适宜浓度VC +BMP2 、IGF1 及bFGF、TGF - β1 均可促进体外传代培养关节软骨细胞的分裂增殖。增殖后的培养细胞仍具有其特异性表型表达 ,而IGF1 培养的细胞则有向成骨样细胞分化的趋向。其综合效能依次为 :VC +BMP2 >bFGF >TGF - β1 >IGF1 。 结论 一定浓度VC +BMP2 及bFGF对适宜密度接种、体外单层传代培养关节软骨细胞具有显著的促增殖作用 ;其中VC +BMP2 具有经济、高效的优点。
Objective To select the high efficiency proliferative agents of optimal accumulation of seeding cells for articular cartilage tissue engineering. Methods The proliferative efficiencies of vitamin C (VC) + bone morphogenetic protein 2 (BMP 2), insulin like growth factor1 (IGF 1) (cultured in 2% serum medium), basic fibroblast growth factor (bFGF) and transforming growth factor β 1 (TGF β 1) (cultured in 10% serum medium) on passage monolayer cultured rabbit articular cartilage cells in an appropriate cencentration cultured in vitro were compared by using 3 H TdR incorporation, MTT intaking and types I and Ⅱ collagen mRNA RT PCR. Results The VC+BMP 2, IGF 1, bFGF and TGF β1 at appropriate concentrations could enhance the division and proliferation of passage monolayer cultured articular cartilage cells in the logarithm growth phase. The proliferated cultured cells still kept specific phenotype as normal articular cartilage cells. But the cells treated with IGF 1 at certain concentrations had the tendency to osteoblast like differentiation. Their proliferative efficiencies were VC+BMP 2> bFGF> TGF β1>IGF 1. Conclusions The VC+BMP 2 and bFGF at appropriate concentrations can significantly enhance the division and proliferation of passage monolayer cultured articular cartilage cells. Among them, the VC+BMP 2 is an economical and high efficiency proliferative agent for articular cartilage tissue engineering.
出处
《中华创伤杂志》
CAS
CSCD
北大核心
2002年第4期235-238,共4页
Chinese Journal of Trauma
基金
国家自然科学基金重点资助项目 (3 980 10 0 )
全军"十0000五"医药卫生科研基金重点资助项目 (0 1Z0 72 )
