人羊膜上皮细胞移植修复兔臂丛神经损伤

中国组织工程研究 第17卷 第53期 2013–12–31出版

Chinese Journal of Tissue Engineering Research December 31, 2013 Vol.17, No.53

http://

doi:10.3969/j.issn.2095-4344.2013.53.010 [http://]

张洋，刘春霞，王亮，张莉，刘伟，赵伟，李治. 人羊膜上皮细胞移植修复兔臂丛神经损伤[J].中国组织工程研究，2013，17(53):9157-9163.

人羊膜上皮细胞移植修复兔臂丛神经损伤*★

张 洋，刘春霞，王 亮，张 莉，刘 伟，赵 伟，李 治(沈阳医学院附属中心医院，辽宁省沈阳市 110024)

文章亮点：

1 人羊膜上皮细胞是克隆原细胞，与胚胎干细胞一样均是全能干细胞，经培养有分化成3个胚层细胞的潜能，而且还保持神经干细胞的特性。

2 实验特点即在于采用体外培养的人羊膜上皮细胞，进行绿色荧光蛋白标记后移植到新西兰大白兔臂丛神经损伤区，经苏木精-伊红染色及荧光显微镜观察，客观的证实了羊膜上皮细胞对兔臂丛神经损伤的修复作用。 关键词：

器官移植；细胞移植；臂丛神经损伤；人羊膜上皮细胞；绿色荧光蛋白；兔；其他基金 主题词：

臂丛神经炎；羊膜；上皮细胞；绿色荧光蛋白质类 基金资助：

沈阳医学院科研基金资助项目(20111031)*

摘要

背景：目前人们把目光多集中在干细胞修复脊髓损伤方面，而对于干细胞移植治疗周围神经损伤的研究却不多，尤其人羊膜上皮细胞修复治疗周围神经损伤的研究和报道更少。 目的：探讨人羊膜上皮细胞对臂丛神经的修复作用。

方法：应用测力神经根拉钩水平牵拉臂丛神经根的方法制备大白兔臂丛损伤模型，提取人羊膜上皮细胞并扩增培养，扫描电子显微镜下观察人羊膜组织及羊膜上皮细胞的超微结构，人羊膜上皮细胞转染绿色荧光蛋白标记并注射入臂丛神经损伤区，苏木精-伊红染色及荧光显微镜观察羊膜上皮细胞对兔臂丛神经损伤的修复作用。 结果与结论：①分离得到的人羊膜上皮细胞表达上皮细胞标志物角蛋白CK19，不表达间充质细胞标志物波形蛋白，不同程度表达CD29，CD73。扫描电子显微镜观察到人羊膜上皮细胞胞体饱满，连接紧密，细胞状态良好。②人羊膜上皮细胞移植到新西兰大白兔臂丛神经损伤模型后18周，免疫荧光检测到神经损伤区可见有表达绿色荧光蛋白的羊膜上皮细胞。③术后6周实验组大白兔前爪功能开始恢复，术后12，18周实验组大白兔前爪功能得到明显改善，功能评分明显提高，对照组几乎没有恢复。以上结果可见人羊膜上皮细胞参与了臂丛神经损伤的修复。

Human amniotic epithelial cell transplantation for repair of brachial plexus injury in rabbits

Zhang Yang, Liu Chun-xia, Wang Liang, Zhang Li, Liu Wei, Zhao Wei, Li Zhi (Central Hospital Attached to Shenyang Medical College, Shenyang 110024, Liaoning Province, China)

Abstract

BACKGROUND: A great deal of attention has been given to stem cell transplantation for repair of spinal cord injury, while there are few studies concerning stem cell transplantation for repair of peripheral nerve injuries, especially transplantation of human amniotic epithelial cells used to repair peripheral nerve injuries.

OBJECTIVE: To discuss the effects of human amniotic epithelial cells in brachial plexus nerve repairing. METHODS: We established rabbit models of brachial plexus injuries and extracted human amniotic epithelial cells for expanding culture followed by ultrastructural observation of human amniotic membrane tissue and amniotic epithelial cells under a scanning electron microscope. Then, human amniotic epithelial cells were transfected with green fluorescent protein markers and injected into the injury site. Hematoxylin-eosin staining and fluorescence microscope were employed to observe the effects of human amniotic epithelial cells in the repair of brachial plexus injuries in rabbits.

RESULTS AND CONCLUSION: (1) Successfully isolated human amniotic epithelial cells expressed cytokeratin 19 rather than vimentin, and also expressed CD29 and CD73 to different extent. Scanning electron microscope

observation showed human amniotic epithelial cells had filled cell bodies and were closed interconnected in good condition. (2) After 18 weeks of successful modeling, human amniotic epithelial cells labeled with green fluorescent protein were visible in the injury site under the fluorescence microscope. (3) In the experimental group, functional recovery of the rabbit forelegs was found at postoperative 6 weeks and dramatically improved at 12 and 18 weeks after surgery. Functional scores were also significantly increased in the experimental group. However, there was no improvement in the control group. These findings indicate that human amniotic epithelial cells are involved

ISSN 2095-4344 CN 21-1581/R CODEN: ZLKHAH 张洋★，男，1981年生，辽宁省沈阳市人，汉族，2011年中国医科大学毕业，硕士，主治医师，主要从事脊柱外科疾病的诊断及治疗。

zhangymed@http://

通讯作者：李治，主任医师，在读博士，沈阳医学院附属中心医院，辽宁省沈阳市 110024 lz8534@http://

中图分类号:R318 文献标识码:A 文章编号:2095-4344 (2013)53-09157-07

修回日期：2013-11-24 (201311021/M

〃W)

Zhang Yang★, Master, Attending physician, Central Hospital Attached to Shenyang Medical College, Shenyang 110024, Liaoning Province, China

zhangymed@http://

Corresponding author: Li Zhi, Studying for doctorate, Chief physician, Central Hospital Attached to Shenyang Medical College, Shenyang 110024, Liaoning Province, China lz8534@http://

Accepted: 2013-11-24

9157

张洋，等. 人羊膜上皮细胞移植修复兔臂丛神经损伤 www.

http://

in the repair of brachial plexus injury.

Subject headings: brachial plexus neuritis; amnion; epithelial cells; green fluorescent proteins Funding: the Scientific Research Fund of Shenyang Medical College, No. 20111031*

Zhang Y, Liu CX, Wang L, Zhang L, Liu W, Zhao W, Li Z. Human amniotic epithelial cell transplantation for repair of brachial plexus injury in rabbits. Zhongguo Zuzhi Gongcheng Yanjiu. 2013;17(53):9157-9163.

0 引言 Introduction

臂丛神经是由第5至8颈神经和第1胸神经前支组成

的；有时C5接受C4的部分神经纤维，T1接受部分T2的神经纤维，凡是组成臂丛的神经受损伤均称为臂丛神经损伤[1]

，造成臂丛神经损伤的主要原因是车祸、运动、重物 …… 此处隐藏：17856字，全部文档内容请下载后查看。喜欢就下载吧 ……