胰岛素样生长因子-1 对脂肪间充质干细胞增殖的影响

目的：探讨胰岛素样生长因子-1(insulin-like growth factor-1，IGF-1)对脂肪间充质干细胞(adipose-derived stem cells，ADSCs) 增殖的影响。方法：采用密度梯度离心法结合贴壁法分离脂肪间充质干细胞，接种于含体积分数为10%的胎牛血清的DMEM 培 养基中行贴壁培养。流式细胞仪检测ADSCs表面标志物(CD90、CD29、CD31、CD34、CD45)的表达情况，利用成骨、成脂诱导液诱 导ADSCs 向成骨细胞、成脂细胞分化，用碱性磷酸酶、油红O 染色观察。采用终浓度为0、5、10、15、20、30 ng/mL IGF的培养基培 养ADSCs，利用Edu 染色标记ADSCs，分析不同浓度的IGF-1 对ADSCs增殖的影响。结果：流式细胞术显示ADSCs的表型分子 CD90、CD29 呈阳性，CD31、CD45 呈阴性，成骨诱导后碱性磷酸酶染色阳性，成脂诱导后油红O染色可见大量脂滴，表明培养的 ADSCs具有成骨、成脂分化的能力。IGF-1 促进ADSCs 增殖的作用随IGF-1 的作用浓度的增加而增加，并逐渐趋于饱和，在趋于 15 滋g/mL的浓度时达到最大促增殖作用，且随着IGF-1 作用时间的延长其促ADSCs 增殖的作用逐渐增强。结论：本实验成功分 离培养ADSCs，IGF-1 对体外培养的ADSCs 有促进增殖的作用。

Effects of insulin-like growth factor-1 on the proliferation of adipose-derived stemcells

Objective: To observe the effects of insulin-like growth factor-1 (IGF-1) on the proliferation of adipose-derived stem cells.Methods:Adipose-derived stem cells (ADSCs) were isolated and purified by density gradient centrifugation combined with adherent method, which were cultured with DMEM with 10% fetal bovine serum (FBS). Flow cytometry was adopted to analyze the expression of surface markers (CD90, CD29, CD31, CD45). Osteoblasts-induced and adipocytes-induced mediums were used to assess the differentiation of ADSCs. Chemical stain method of alkaline phosphatase and oil red O were used to observe the ADSCs, which were cultured with IGF-1 at 0, 5, 10, 15, 20, 30 ng/ml for 24, 48, 72 hours and labelled with Edu respectively.Results:CD90, CD29 of ADSCs were positively expressed and CD31, CD45 were negatively expressed. Alkaline phosphatase staining was positive. And oil red O staining showed a large number of lipid droplets. ADSCs had the petentiality to differentiate into osteoblasts and adipocytes after inducement. IGF-1 promoted the proliferation of ADSCs in a concentration and time dependent manner, which reached peak at 15 ug/mL.Conclusion:ADSCs of SD rats were isolated and cultured sucessfully, IGF-1 had promotion effect on the proliferation of ADSCs in vitro.