摘要
目的重编程诱导大鼠骨髓间充质干细胞(MSCs)向胰岛内分泌细胞分化并探讨其可能的分化机制。方法原代培养的大鼠骨髓MSCs经分离、培养至第3代后,用负向调节因子Rest/Nrsf、Shh慢病毒干扰载体转染并培养10 d,通过实时定量RTPCR和免疫荧光检测胰岛细胞分化相关基因的表达。结果 MSCs慢病毒转染后,胰岛素、胰高血糖素及生长抑素的表达明显升高,并表达胰岛内分泌细胞分化的相关基因。结论通过转染shRest/Nrsf、shShh慢病毒载体能够诱导大鼠MSCs向胰岛内分泌细胞分化,EGFR、ERK1、MMP以及PI3K等参与这一分化过程。
Objective To induce insulin-producing cells from rat bone marrow- derived mesenchymal stem cells （MSCs）in vitro by suppressing two repressor genes repressor element- 1 silencing transcription factor/neuron- restrictive silencer factor （Rest/Nrsf） and sonic hedgehog （Shh）, and discuss the possible mechanism. Methods Rat bone marrow-derived MSCs were separated and cultured to the third generation, and then tmnsfected with the shRest/Nrsf and shShh lentivirus vectors and cultured for another ten days. Real-time quantitative RT-PCR and immunofluorescent assay were employed to detect related gene expression of islet cell differentiation. Results Insulin, glueagon and somatostatin increased significantly after lentivims transfection, and pancreas islet endocrine cell differentiation related genes were expressed. Conclusion Rat MSCs can be induced to islet endocrine cell differentiation by eolxansfeetion of shRest/Nrsf and shShh lentivirus vectors. ERK1, MMP, EGFR, PI3K are involved in this process of differentiation.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2016年第1期1-6,共6页
Journal of China Medical University
基金
国家自然科学基金(81370883)
国家基础研究资助项目(2012CB518103)
教育部博士基金(20132104110020)
辽宁省科技厅博士启动基金(2014010534-301)
