生长分化因子5促进脂肪干细胞成软骨分化的实验研究

目的研究生长分化因子（GDF5）在诱导脂肪干细胞（ADSCs）成软骨分化中的作用。方法大鼠ADSCs在不同诱导条件下经细胞微团培养后,通过检测细胞增殖、糖胺聚糖和蛋白聚糖、软骨细胞特异性基因CollagenⅡ、AggrecanmRNA和蛋白质表达,以及软骨细胞肥大标志基因CollagenⅠ、CollagenⅩ的表达水平,比较不同浓度的GDF5（10ng/ml、100ng/ml和1000ng/ml）与TGFβ1（10ng/ml）对ADSCs成软骨分化的促进作用。结果 GDF5对AD-SCs成软骨分化过程中的细胞增殖具有显著的促进作用,同时还能增加细胞外基质糖胺聚糖和蛋白聚糖的含量,以及上调软骨细胞特异性CollagenⅡ和Aggrecan基因和蛋白表达。在10ng/ml、100ng/ml和1000ng/ml的GDF5剂量组中以100ng/ml为最佳浓度,其效果显著优于10ng/ml的TGFβ1。结论本研究结果表明GDF5能够显著促进脂肪干细胞的成软骨分化,并且以100ng/ml的效果最为明显。

Role of growth and differentiation factor 5 on chondrogenesis of adipose derived stem cells

Objective To study the effects of GDF5 on promoting chondrogenesis of ADSCs.Methods Cell proliferation,glycosaminoglycan and proteoglycan,the mRNA and protein expression of chondrocyte marker genes collagen Ⅱ and aggrecan,and the expression of the marker genes of chondrocyte hypertrophy collagen Ⅰ and collagen Ⅹ were determined in the ADSCs under different induce conditions in micromass culture system.The promoting chondrogenic effects of 10ng/ml,100ng/ml,1000ng/ml GDF5 were compared with those of 10ng/ml TGFβ1.Results GDF5 could promote the proliferation of ADSCs cells in the process of chondrogenesis,increase the contents of extracellular matrix glycosaminoglycan and proteoglycan,and up-regulate the genomic and proteinic of chondrocyte marker genes collagen II.100ng/ml GDF5 was the optimal concentration among 10ng/ml,100ng/ml and 1000ng/ml groups.Conclusion The results of this study demonstrates that GDF5 could promote ADSCs chondrogenesis significantly,and 100ng/ml GDF5 was the optimal concentration.