糖稳态调节细胞定向分化的基因改造方案探讨

To explore the gene modified strategy for stem cells directional differentiation by the glucose-steadied regulation

目的:探索糖稳态调节细胞定向分化的基因改造方案思路。方法:①研究构建的解除干细胞分化抑制状态的信号传导与转录激活因子-3(STAT-3)基因载体,检测其介导目的基因感染AR IP等细胞基因表达;②研究构建的活化配体DL诱导N信号、在“旁侧抑制”过程中扮演重要角色的Kuzban ian(KUZ)基因载体在转基因小鼠表达后糖稳态调节细胞的分化。结果:在完成的STAT-3腺病毒基因载体生产了腺病毒,用带有目的基因的腺病毒对AR IP靶细胞进行感染,3 d后,荧光显微镜观察时,活细胞、固定液固定细胞均看到了构建基因中绿色荧光蛋白成功、高效的阳性表达报告。在完成的Kuzban ian(KUZ)基因载体构建、转基因、动物模型、小鼠品系鉴定后,用非放射性原位杂交实验对胰腺靶细胞的mRNA进行检测,获得了预期的阳性结果。结论:通过用某些分化抑制性基因的显性失活形式(DN)竞争性预先解除干细胞分化的抑制状态,然后再定向分化诱导使之朝向期望的目标细胞方向分化,可能是糖稳态细胞定向分化的可行途径之一。

Objective：To investigate the strategy of gene modified for stem cells directional differentiation by the glucosesteadied regulation. Methods： 1. The expression of STAT-3 （ signal transducer and activators of transcription, STATs） protein and cells differentiation in the ARIP etc. cells were studied by gene modified and differentiation induced; 2. Cells differentiation was studied by the expression of KUZ mRNA in the mouse pancreas islet cells after differentiation inhibited. Results：There was strong positive of immunofluorescence in ARIP cells after infected by Adenovirus assisted STAT-3 gene. Non-radioactivity in situ hybridization studies on the expression of Kuzbanian（KUZ） gene in trans-gene mice pancreas islet were positive. Conclusion： Gene modified induced by factor differentiation may be an availability approach of stem cells directional differentiation about the glucose-steadied regulation.