解剖学杂志2012，Vol.35Issue(2)：149-153,5.
维甲酸联合音猬因子诱导人脐带间充质干细胞分化为运动神经元
Differentiation of human umbilical cord mesenchymal stem cells into motoneurons induced by retinoic acid combined with sonic hedgehog
摘要
Abstract
Objective :To explore the feasibility that human umbilical cord mesenchymal stem cells (hUCMSCs) differentiated into motoneurons induced by retinoic acid (RA) and sonic hedgehog (Shh), thereafter, to provide experimental basis for clinical translation. Methods-, The hUCMSCs were isolated and purified by double enzymatic digestion and the surface antigens of these cells were detected by flow cytometry. According to different inducer. the experimentation was divided into four groups; control (no inducer), RA group (0. 5 μg/ml RA), Shh group (300 ng/ml Shh) and RA+Shh group (0. 5 μg/ ml RA+300 ng/ml Shh). After hUCMSCs being induced for 25 days, the morphological change was observed, and the expressions of Nestin, MAP-2, Hb9 and GFAP were detected by immunocytochemical method. Additionally, the immunocy-tochemical analysis for these four proteins was also performed on day 5, 10, 15, and 20 after induction in RA+Shh group. Results-. hUCMSCs were positive to CD29 and CD44, and negative to CD31 and CD34. In the control and Shh group, morphological change of cells was not found, and the proportion of positive cells of Nestin and MAP-2 was low. Huwever, the cells induced by RA only or combination of RA+Shh presented typical neuronal-like appearance exhibited as cytoplasm contracted towards the nucleus and processes were formed and connected like network. The proportion of positive cells of Nestin and MAP-2 was significantly higher than that in the other groups. In four groups, positive to Hb9 was only found in RA+ Shh group. And, the GFAP was weak expressed without significant difference in four groups. The combination of RA and Shh caused detectable Nestin on day 5, MAP-2 on day 10, Hb9 on day 15. On day 20, the Nestin expression was decreased, and MAP-2 and Hb9 reached their highest levels on day 25. Conclusion-. RA could greatly promote the differentiation of hUCMSCs into neurons, while solitary Shh have no effect on it. However, hUCMSCs can be induced into the motoneurons with combination of RA and Shh.关键词
人脐带间充质干细胞/运动神经元/维甲酸/音猬因子/诱导Key words
human umbilical cord mesenchyrmal stem cells/ mo-toneurons/ retinoic acid/ sonic hedgehog/induction引用本文复制引用
刘学元,李德华,单伟,姜东,房艳..维甲酸联合音猬因子诱导人脐带间充质干细胞分化为运动神经元[J].解剖学杂志,2012,35(2):149-153,5.基金项目
辽宁省自然科学基金(20102138) （20102138）
辽宁医学院青年科技启动基金(Y2010Z004) （Y2010Z004）
