李佳丹 陈婷 田洪榛 陈江 陈光辉.胰岛素样生长因子-1 对脂肪间充质干细胞增殖的影响[J].,2014,14(14):2615-2621 |
胰岛素样生长因子-1 对脂肪间充质干细胞增殖的影响 |
Effects of insulin-like growth factor-1 on the proliferationof adipose-derived stemcells |
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DOI: |
中文关键词: 胰岛素样生长因子-1 脂肪间充质干细胞 增殖 |
英文关键词: Insulin-like growth factor-1 Adipose-derived stemcells Proliferation |
基金项目:国家高技术研究发展计划(2011AA020101) |
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中文摘要: |
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。 |
英文摘要: |
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. |
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