中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (32): 5951-5956.doi: 10.3969/j.issn.1673-8225.2011.32.014

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

诱导人眼轮匝肌来源肌源干细胞向许旺细胞样细胞的分化

丁维进1，2，张文俊1，2，孙美庆1，2，苏志达2，李  翠2，刘安堂1，2，江  华1，2   

  1. 解放军第二军医大学长征医院，1  整形外科，2神经科学研究中心，上海市 200003
  • 收稿日期:2011-04-23 修回日期:2011-06-15 出版日期:2011-08-06 发布日期:2011-08-06
  • 通讯作者: 江华，博士，教授，主任医师，解放军第二军医大学长征医院，整形外科，神经科学研究中心，上海市 200003 docdwj@163.com
  • 作者简介:丁维进☆，男，1982年生，江苏省海安县人，汉族，2011年解放军第二军医大学毕业，博士，主要从事周围神经损伤后修复研究。 dingweijin@yahoo.cn
  • 基金资助:

    上海市基础研究重点课题(08JC1407100)。

Differentiation potential of human Orbicularis oculi muscle-derived stem cells towards Schwann cells phenotype

Ding Wei-jin1,2, Zhang Wen-jun1,2, Sun Mei-qing1,2, Su Zhi-da2, Li Cui2, Liu An-tang1,2, Jiang Hua1,2   

  1. 1Plastic and Reconstructive Department of Changzheng Hospital, Second Military Medical University of Chinese PLA, Shanghai  200003, China; 2Neuroscience Research Center of Changzheng Hospital, Second Military Medical University, Shanghai  200433, China
  • Received:2011-04-23 Revised:2011-06-15 Online:2011-08-06 Published:2011-08-06
  • Contact: Jiang Hua, Ph.D., Professor, Chief physician, Plastic and Reconstructive Department of Changzheng Hospital, Second Military Medical University of Chinese PLA, Shanghai 200003, China; Neuroscience Research Center of Changzheng Hospital, Second Military Medical University, Shanghai 200433, China docdwj@163.com
  • About author:Ding Wei-jin☆, Doctor, Plastic and Reconstructive Department of Changzheng Hospital, Second Military Medical University of Chinese PLA, Shanghai 200003, China; Neuroscience Research Center of Changzheng Hospital, Second Military Medical University, Shanghai 200433, China dingweijin@yahoo.cn
  • Supported by:

    the Key Basic Research Project of Shanghai, No. 08JC1407100*

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被引次数

1

摘要：

背景：肌源干细胞的优越性引导学者们尝试从人眼轮匝肌中分离该细胞，同时进行许旺细胞方向的诱导分化，为周围神经的修复提供新的种子细胞来源。
目的：诱导人肌源干细胞向具有许旺细胞特性的细胞分化。
方法：①收集重睑成形术中切除的上睑眼轮匝肌，经酶消化和细胞筛过滤，贴壁培养分离人肌源干细胞，予细胞特异标记物以免疫组织化学染色。②取大鼠坐骨神经分离培养许旺细胞，收集许旺细胞条件培养液。③将人肌源干细胞与许旺细胞条件培养液共培养，观察转化细胞的形态和免疫组织化学染色的变化。
结果与结论：①原代培养4周时可见人肌源干细胞，与传代培养之人肌源干细胞同样呈现明显的小圆形形态，折光性强，少量细胞呈现短梭形。所有人肌源干细胞表现为Desmin阳性，Sca-1染色阳性。②分离培养大鼠坐骨神经来源许旺细胞，S100染色阳性率为(97.4±0.7)%。③经与许旺细胞条件培养液共培养，人肌源干细胞分化后细胞表达许旺细胞特异标记物S100，GFAP和p75。结果提示分离获得人肌源干细胞与许旺细胞条件培养液共培养，可以诱导人肌源干细胞表达许旺细胞特异标记物，初步证实了人肌源干细胞可分化为许旺细胞样细胞。

关键词: 人肌源干细胞, 眼轮匝肌, 重睑成形术, 分化, 许旺细胞, 周围神经损伤, 再生医学

Abstract:

BACKGROUND: Muscle derived stem cells (MDSCs) can be isolated from human orbicularis oculi muscle and be differentiated to a Schwann cell phenotype which could eventually provide functional benefits for peripheral nerve repair.
OBJECTIVE: To induce the differentiation of MDSCs into Schwann cell phenotype.
METHODS: ①Under the support of microscope, we collected the discarded human Orbicularis oculi muscle resected in the upper eyelid blepharoplasty and isolate human-MDSCs within it with aid of tri-enzyme digestion and pre-plating technique, and then identify the cells by immunohistochemistry method. ②We isolated Schwann cells and identify the cells by immunohistochemistry method. Through half-harvest method, we would like to prepare conditioned medium from Schwann cell culture. ③We co-culture human-MDSCs with Schwann cell conditioned medium and the transdifferentiated cell morphology was investigated daily under microscope. The common used marker, S-100, GFAP and p75 were stained to identify Schwann cell phenotype with use of immunohistochemistry method.
RESULTS AND CONCLUSION: ①We collected human Orbicularis oculi muscle sample from three young female volunteer with their consensus. Human-MDSCs were isolated from Orbicularis oculi muscle and have their desmin positively stained and Sca-1 was positively expressed. ②Schwann cells were isolated and identified with S-100 positively stained at the rate of (97.4±0.7)%. ③The isolated human-MDSCs were successfully transdifferentiated into Schwann cell-like cells with positive expression of S100, GFAP and p75, which would serve as a unanimous evidence of Schwann cell phenotype. Human-MDSCs could be transdifferentiated into Schwann cell-like cells when co-cultured within Schwann cell conditioned medium, which would serve as an alternative candidate for commonly studied Schwann cells in tissue engineering nerve graft.

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