不同微环境对骨髓间充质干细胞分化为产胰岛素细胞的影响
The impact of different microenvironments on the differentiation of rat bone marrow mesenchymal stem cells into insulin-producing cells
摘要 目的 观察不同的微环境对大鼠骨髓间充质干细胞(MSCs)体外分化成为产胰岛素细胞(IPCs)的影响.方法 采用贴壁法分离纯化大鼠MSCs,用不同的微环境进行诱导,对照组诱导剂为含有角朊细胞生长因子、ITS、尼克酰胺的无血清DMEM/F12培养基、实验组在对照组基础上添加胰腺条件培养液;对诱导后细胞进行光、电镜观察,双硫腙和免疫细胞化学进行鉴定,并进行体外葡萄糖刺激实验,测定细胞分泌胰岛素及C-肽功能.结果 实验组及对照组均可诱导分化为IPCs,但实验组分化而成的IPCs在数量及功能上均高于对照组.结论 大鼠胰腺提取物模拟的微环境能促进大鼠MSCs体外诱导分化为较高质量的IPCs.
Abstract:
Objective To study the impact of different microenvironment on difierentiation of rat bone marrow mesenchymal stem cells(MSCs)into insulin-producing cells(IPCs)in vitro.Methods MSCs were isolated from rat bone marrow and purified by their adhesive process onto the culture dish.The descended cells were then cultivated in vitro mimicking different microenvironments,in which control group was cultivated in serllm free DMEM/F12 medium including keratinocyte growth factor(KGF),ITS and Nicotinamide,while experiment group was cultivated in sermn free DMEM/F12 medium in which the rat pancreatic extract wag added.During the cultivation period the cells were taken for light and electron microscopy at different time points,identified by dithizone(DTZ)and immunocytochemistry.The glucose stimulation test was performed in vitro and the levels of iasulin and C-peptide in response to the difierent glucose concentrations were assayed.Results The MSCs could difierentiate into IPCs both in the control group and the experimental group.The IPCs harvested from experiment group had greater number and better function than that from control group.Conclusion The mieroenvironment in which the rat pancreatie extract was added could promote the differentiation of rat bone marrow MSCs into better IPCS in vitro.
作者单位: 第四军医大学西京医院肝胆外科,西安,710032
刊 名 中华实验外科杂志 ISTICPKU
年,卷(期): 2008, 25(3)
分类号: R3
机标分类号: R73 R58
在线出版日期: 2008年5月5日