基础医学与临床 ›› 2009, Vol. 29 ›› Issue (1): 47-51.

• 研究论文 • 上一篇    下一篇

大鼠肌源性干细胞体外转分化为胰岛素分泌细胞

刘畅 刘福强 梅晰凡 杨彪   

  1. 辽宁医学院 第一附属医院 辽宁医学院 第一附属医院 辽宁医学院 第一附属医院 辽宁医学院 第一附属医院
  • 收稿日期:2007-10-08 修回日期:2008-06-02 出版日期:2009-01-25 发布日期:2009-01-25
  • 通讯作者: 刘畅

Transdifferentiation of Rat Muscle Derived Stem Cells into Insulin-Secreting Cells in Vitro

Chang LIU, Fu-qiang LIU, Xi-fan MEI, Biao YANG   

  1. The First Afiliated Hospital, Liaoning Medical College The First Afiliated Hospital, Liaoning Medical College The First Afiliated Hospital, Liaoning Medical College The First Afiliated Hospital, Liaoning Medical College
  • Received:2007-10-08 Revised:2008-06-02 Online:2009-01-25 Published:2009-01-25
  • Contact: Chang LIU,

摘要: 目的 探讨体外自体肌源性干细胞(MDSCs)定向诱导分化为胰岛素分泌细胞团的可行性。方法 用差速贴壁的方法对SD大鼠MDSCs进行分离和培养,加入联合诱导剂培养7~12d。通过形态学观察、双硫腙(DTZ)染色、免疫细胞化学染色、葡萄糖刺激实验和RT-PCR,对诱导细胞进行体外结构和功能鉴定。 结果 联合诱导后12d有典型的胰岛样结构出现;诱导后第12天细胞团表达胰岛素,而未诱导组细胞不表达;诱导后第7天检测到胰岛素 (INS)、胰高血糖素(GLU)和生长抑素(SS)基因的表达。 结论 体外诱导大鼠肌源性干细胞可定向分化为胰岛素分泌细胞存在可行性,本实验将可能为临床糖尿病干细胞治疗提供新的途径。

关键词: 肌源性干细胞, 转分化, 尼克酰胺, 胰岛素, 肌源性干细胞, 转分化, 尼克酰胺, 胰岛素

Abstract: Objective To explore the possibility of diferentiating functional insulin-secreting cells from muscle derived stem cells.Methods SD rat muscle derived stem cells (MDSCs)were isolated and cultured then induced for 7~12 days with combination inductor.Morphological changes of the induced cells were observed under an inveted microscope.DTZ staining ,immunocytochemical staining were conducted to examine the production of insulin. Glucose challenge and radio immunoassay were perfomed to evaluate the function of the aggregates,and RT-PCR was conducted to detect the expression of endocrine gene.Results Typical insulin-producing cells were observed at the end of the protocol(12 days). Ditizone staining and immunohistochemistry for insulin were both positive. Insulin was secreted by these cells in response to different concentrations of glucose stimulation in a regulated manner (P<0.05). RT-PCR showed the insulin,glucagon and SS gene expression on the 7th day after the inducement of combination inductor. Conclusion Insulin-secreting cells can be transdiferentiated from muscle derived stem cells,which may be a new procedure for clinical treatment of diabetes.

Key words: Muscle-derived stem cells, Transdiferentiated, Nicotinamide, Insulin., muscle-derived stem cells, transdiferentiated, nicotinamide, insulin