摘要
目的探索大鼠骨髓间质干细胞(MSCs)体外诱导分化为胰岛素分泌细胞的方法,为解决胰岛移植物来源匮乏问题提供新的思路。方法采用横向分化技术,将成年大鼠MSCs诱导成胰岛素分泌细胞;间接免疫荧光法鉴定诱导前后的细胞巢蛋白、胰岛素、胰高血糖素、生长抑素的表达;RTPCR法检测诱导前后的细胞胰岛素1、Pdx1、Pax6mRNA的表达;24h累积分泌量测定和胰岛素刺激实验评价诱导前后细胞的功能。结果诱导5h,巢蛋白阳性细胞为(44.6±7.3)%,诱导24h增至(61.8±8.4)%。此后,巢蛋白阳性细胞数开始下降,诱导第14天后,巢蛋白表达基本消失。而且,诱导后细胞可表达胰岛素、胰高血糖素、生长抑素等蛋白;表达胰岛素1及其多种转录因子基因mRNA;胰岛素刺激实验反应敏感,而诱导前MSCs不具备上述特点。结论体外大鼠MSCs可诱导成为胰岛素分泌细胞,为胰岛移植开辟新的研究思路。
Objective To induce bone marrow mesenchymal stem cells （MSCs） to differentiate into insulin-secreting cells in vitro. Methods Using a defined culture medium and technique for transdifferentiation, MSCs from adult SD rats were guided into specific insulin-secreting cells. The expressions of islet-specific hormones and proteins, such as insulin, glucagons and somatostatin were detected and analyzed by indirect immunofluorescence cytochemistry staining before and after the induction. And insulin 1, Pdx-1 and Pax-6 were detected by reverse transcription-PCR （RT-PCR）. In addition, insulin secretion was examined using radioimmunoassay. Results The differentiated MSCs which expressed nestin were 44.6±7.3% after 5 h induction, 6. 18±8.4% after 24 h induction and disappeared at 14 day culture. Meantime, the differentiated cells had positive immunoreactivity to insulin, glucagon and somatostatin,and expressed insulin 1, Pdx-1, Pax-6 mRNA. The differentiated cells were able to produce higher level insulin, and displayed glucose-dependent insulin release in vitro. Conclusion Adult rat MSCs can be induced to differentiate into insulin-secreting cells in vitro.
出处
《中国糖尿病杂志》
CAS
CSCD
北大核心
2006年第2期126-128,共3页
Chinese Journal of Diabetes
基金
国家自然科学基金资助项目(30200128)
关键词
干细胞
骨髓
胰岛
分化
Stem cells, myeloid
Islet of Langerhans
Differentiation
