大鼠骨髓基质干细胞分化为胰岛样细胞  

Islet-like cells differentiated from rat marrow mesenchymal stem cells

作　　者：陈立波[1] 姜小兵[1] 杨炼[1] 

机构地区：[1]华中科技大学协和医院外科,湖北省武汉市430022

出　　处：《世界华人消化杂志》2004年第4期871-874,共4页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30170911湖北省科技攻关计划;No.2002AA301C26~~

摘　　要：目的:探讨体外自体骨髓基质干细胞(MSC)定向诱导分化为胰岛样细胞的可能性,为进一步诱导分化胰岛细胞治疗糖尿病奠定实验和理论基础. 方法:对Wistar 大鼠骨髓基质干细胞(MSC)进行体外培养,通过不同的条件诱导MSC向胰岛样细胞分化:LN 组,10 mmoL/L尼克酰胺+1 mmoL/Lβ-巯基乙醇的L-DMEM(含20 mL/L FBS)预先诱导24h,10mmoL/L 尼克酰胺+lmmol/Lβ-巯基乙醇的无血清H-DMEM诱导10h.HN组,含20 mmoL/L尼克酰胺的L-DMEM (20 mL/L FBS)预先诱导24 h,20 mmoL/L尼克酰胺的无血清H-DMEM诱导10h.诱导细胞形态通过倒置显微镜观察,并通过免疫细胞化学检测分化细胞表达Insulin 和Nestin.向3只Streptozotocin(STZ)诱导糖尿病大鼠皮下种植MSC分化胰岛细胞1×107,1 wk后观察大鼠血糖水平变化. 结果:LN、HN组细胞均可见部分骨髓基质干细胞分化、增生,形成胰岛样细胞团,部分细胞向神经元样细胞分化,对照组细胞未见明显分化.免疫细胞化学显示Insulin在诱导细胞呈显著表达,Nestin在分化前细胞显著表达.皮下种植MSC分化胰岛细胞可有效降低STZ 糖尿病大鼠的血糖水平. 结论:体外诱导骨髓基质干细胞定向分化为胰岛样细胞存在可行性,本实验将可能为临床糖尿病干细胞治疗提供新的途径.AIM: To explore the possibility of differentiating functional islet-like cells from marrow mesenchymal stem cells. METHODS: Rat marrow mesenchymal stem cells (MSC) were isolated from Wistar rats and cultured. Passaged MSCs were induced to differentiate into islet-like cells under following condition: LN group, pre-induced with L-DMEM including 10 mmoL/LNicotinamide+1 mmoL/L β-Mercapto-ethanol +20 mL/L fetal bovine serum for 24 h,then induced with serum free H-DMEM solution including 10 mmoL/L nicotinamide +1 mmoL/L β-mercaptoethanol for 10h. HN group, pre-induced with L-DMEM including 20 mmoL/L Nicotinamide +20 mL/L fetal bovine serum for 24 h, then induced with serum free H-DMEM solution including 10 mmoL/L nicotinamide for 10 h. Differentiated cells were observed under inverse microscope, insulin and nestin expressed in differentiated cells were detected with immunocytochemistry. Diabetes model was induced with streptozotocin (STZ) injection, and 3 diabetes rats were subcutaneously injected with 1×107 islet cells differentiated from MSC respectively, and blood glucose level was tested 1 week after cell injection. RESULTS: Islet -like clustered cells were observed under both LN and HN groups, abundant insulin expression was detected in islet-like cells, and nestin was detected in pre-differentiated cells. Glucose level in STZ-diabetic rats could be effectively controlled by islet cells differentiated from MSC. CONCLUSION: Islet-like functional cells can be differentiated from marrow mesenchymal stem cells, which may be a new procedure for clinical treatment of diabetes.

关 键 词：大鼠 骨髓基质干细胞 细胞分化 胰岛样细胞 MSC 糖尿病 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]