[1]胡琳燕,余加林,李芳,等.丹参诱导骨髓间充质干细胞分化为神经样细胞的突触功能研究[J].第三军医大学学报,2009,31(02):144-147.
 HU Lin-yan,YU Jia-lin,LI Fang,et al.Synapse function of neuron-like cells induced from mesenchymal stem cells by Salvia miltiorrhiza[J].J Third Mil Med Univ,2009,31(02):144-147.
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丹参诱导骨髓间充质干细胞分化为神经样细胞的突触功能研究(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
31卷
期数:
2009年第02期
页码:
144-147
栏目:
论著
出版日期:
2009-01-30

文章信息/Info

Title:
Synapse function of neuron-like cells induced from mesenchymal stem cells by Salvia miltiorrhiza
作者:
胡琳燕余加林李芳刘官信李禄全
重庆医科大学附属儿童医院新生儿科
Author(s):
HU Lin-yan YU Jia-lin LI Fang LIU Guan-xin LI Lu-quan
Department of Neonatology, Children’s Hospital, Chongqing Medical University, Chongqing 400014, China
关键词:
MSCs丹参神经性分化突触功能
Keywords:
mesenchymal stem cells Salvia miltiorrhiza neuron differentiation synapse function
分类号:
R282.71;R329.21;R338.13
文献标志码:
A
摘要:
目的    探讨由丹参诱导骨髓间充质干细胞(mesenchymal stem cells, MSCs)定向分化的神经细胞是否具有突触循环功能。    方法    采用优化诱导方案对4~5代状态好的MSCs进行反复多次诱导,终止诱导后分别进行免疫荧光细胞化学、细胞内钙离子流及突触功能检测。以50 mmol/L KCl作为诱发动作电位的刺激因子,采用激光共聚焦显微镜 (LSCM) 对Fluo-3/AM和SynaptoRed C-2荧光探针进行荧光激发,检测分化神经细胞在细胞外高钾刺激下细胞内钙流及细胞突触功能。    结果    4次诱导5 h后,神经样细胞伸出突起并交互成复杂网状;免疫荧光细胞化学显示4次诱导5 h后TUJ-1表达率为(96.7±2.8)%,突触小泡蛋白(synaptophysin)表达率为(96.2±2.1)%;LSCM显示4次诱导5 h后细胞在高钾刺激下细胞内钙流增加,细胞突触发生胞吞胞吐。    结论    丹参可高效快速诱导MSCs分化为神经细胞,此细胞具有正常的突触循环功能。
Abstract:
Objective    To explore whether the neurons induced from mesenchymal stem cells (MSCs) have synapse function or not.     Methods    Passage 4-5 MSCs in good shape was induced by Salvia miltiorrhiza with optimized protocol for several times, and then the observation under inverted phase contrast microscope, immunofluorocytochemistry and the measurement of neurophysiological function were carried out. Ca2+ influx and synapse function were detected with laser-scanning confocal microscopy(LSCM), taking 50 mmol/L KCl as stimuli to evoke action potential.     Results    The cells that had been induced for 5 h at 4th time from MSCs, looked like neurons and displayed that the processes stretched out to form complex net. Immunofluorocytochemistry presented that the rate of TUJ-1 expression was (96.7±2.8)% and that of synaptophysin was (96.2±2.1)%. When the neuron-like cells were stimulated by high concentration of KCl, intracellular Ca2+ influx enhanced quickly. When the neuron-like cells were stimulated by high KCl solution for the first time, SynaptoRed-C2 anchored onto the membrane, and after the second excitation, the fluorescence intensity decreased quickly.     Conclusion    The neuron-like cells derived from MSCs that are induced by salvia miltiorrhiza with optimized protocol have synapse function.

参考文献/References:

胡琳燕, 余加林, 李芳, 等. 丹参诱导骨髓间充质干细胞分化为神经样细胞的突触功能研究[J]. 第三军医大学学报, 2009, 31(2):144-147.

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更新日期/Last Update: 2009-01-08