大鼠皮肤成纤维细胞分泌的细胞外基质对肌腱干细胞特性的影响

The effects of extracellular matrix generated from rat dermal fibroblasts on the characteristics of tendon stem cells

[目的]探讨皮肤成纤维细胞分泌的细胞外基质（ECM）对肌腱干细胞增殖及腱向分化的影响。[方法]选取雄性SD大鼠10只,体重285~310 g,无菌切取皮肤及髌腱组织,进行皮肤成纤维细胞及肌腱干细胞的分离和培养。制备成纤维细胞分泌的ECM包被的培养皿,以空白培养皿为参照,在肌腱干细胞接种于培养皿后,计算细胞倍增时间,利用细胞免疫荧光检测干细胞表面标志。利用RT-PCR检测腱向分化相关基因的表达。[结果]成纤维细胞分泌的ECM显著缩短了肌腱干细胞的倍增时间（P〈0.01）。成纤维细胞分泌的ECM能明显诱导肌腱干细胞分化,同时其可以有效促进腱向分化相关基因的表达。[结论]皮肤成纤维细胞分泌的ECM可以有效促进肌腱干细胞增殖及腱向分化,这为优化肌腱干细胞的实验研究提供了有利的理论依据,为肌腱损伤后优质愈合提供了新的思路方法。

[ Objective] To determine the effects of extracellular matrix （ECM） generated from rat dermal fibroblasts on the proliferation and tenogenic differentiation of tendon stem cells （TSCs） . [ Methods] Ten adult male SD rats （weighing between 285 and 310 g） were used for this study. The dermis and patellar tendons of the rats were sterilized and isolated. This was followed by the isolation and culture of dermal fibroblast and patellar TSCs. The TSCs were then divided into 2 groups and cul- tured in plates coated with extracellular matrices secreted by dermal fibroblasts or in uncoated plates. Proliferative capacity and characterization of TSCs cultured in dFM were determined by population doubling time, immunofluorescence staining, and qRT -PCR. [ Results ] The ECM produced by dermal fibroblasts promoted the proliferation and differentiation of TSCs in vitro. Moreover, tendon- specific -gene -expression was significantly improved by ECM. [ Conclusion ] ECM produced by dermal fibroblasts induced TSC proliferation and tenogenic differentiation. These findings provide a theoretical basis for the optimization of TSC proliferation and tendon repair.