地黄低聚糖对人脂肪组织来源干细胞旁分泌作用的影响

摘要

目的 探讨地黄低聚糖(RGOs)对分离、培养的人脂肪组织来源干细胞(hASCs)旁分泌作用的影响.方法 酶消化法及贴壁法从人腹部脂肪组织中分离、培养hASCs.流式细胞仪检测hASCs表面抗原CD29、CD31、CD44、CD45、CD90、CD105、HLA-DR的表达.hASCs经RGOs(0.1 g/L)预处理12 h并继续干预,ELISA法观察不同时间点hASCs培养上清液中血管内皮生长因子(VEGF),肝细胞生长因子(HGF),胰岛素样生长因子-1(IGF-1),碱性成纤维细胞生长因子(bFGF),基质细胞衍生因子-lα(SDF-1α)浓度的变化.结果 hASCs表面抗原CD29、CD44、CD90、CD105表达呈阳性,CD31、CD45、HLA-DR表达呈阴性.与对照组相比,RGOs可以促进体外培养的hASCs分泌VEGF、HGF、IGF-1、SDF-1 α(P ＜0.05),而对于bFGF的分泌无明显影响.结论 RGOs可以增强体外培养的hASCs的旁分泌作用.%ObjectiveTo isolate and culture human adipose tissue-derived stem cells (hASCs) in vitro and investigate the effect of Rehmannia glutinosa oligosaccharides (RGOs) on its paracrine action. Methods The isolation and culture of hASCs was conducted by enzyme digestion and adherent from adipose tissue from abdomen. The surface antigens (CD29.CD31 ,CD44,CD45 ,CD90,CD105 and HLA-DR) of hASCs were examined by flow cytome-try. The levels of vascular endothelial growth factor (VEGF) , hepatocyte growth factor ( HGF), insulin-like growth factor-1 (IGF-1) ,basic fibroblast growth factor (bFGF) and stromal cell derived factor-la (SDF-la) in the hASCs supernatant were determined by enzyme-linked immunosorbent assay (ELISA) after RGOs (0. 1 g/L) pretreating for 12 hours. Results The hASCs were identified by positive expression of CD29, CD44, CD90 and CD105, and negative expression of CD31 ,CD45 and HLA-DR. Campared with control group,the secretion of VEGF,HGF,IGF-1 and SDF-lot in RGOs group was increased significantly(P ＜ 0. 05) ,but the secretion of bFGF did not increase significantly. Conclusion RGOs could enhance the paracrine action in cultured hASCs in vitro.