中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (25): 4591-4595.doi: 10.3969/j.issn.1673-8225.2012.25.007

• 复合支架材料 composite scaffold materials • 上一篇    下一篇

NgR基因沉默骨髓间充质干细胞/许旺细胞支架复合体的构建

王 东，张建军，杨卫山   

  1. 天津市第四中心医院神经外科，天津市 300140
  • 收稿日期:2011-12-03 修回日期:2012-02-03 出版日期:2012-06-17 发布日期:2013-11-04
  • 作者简介:王东★，男，1971年生，河北省乐亭县人，汉族，2011年天津医科大学毕业，硕士，副主任医师，主要从事脊髓损伤研究。 wd5609@ hotmail.com
  • 基金资助:

    天津市卫生局科技基金面上课题(2010ky04)；天津市应用基础研究计划面上项目(12JCYBJC18000)

Construction of the scaffold composite of bone marrow mesenchymal stem cells and Schwann cells after NgR gene silencing

Wang Dong, Zhang Jian-jun, Yang Wei-shan   

  1. Department of Neurosurgery, Tianjin Fourth Central Hospital, Tianjin 300140, China
  • Received:2011-12-03 Revised:2012-02-03 Online:2012-06-17 Published:2013-11-04
  • About author:Wang Dong★, Master, Associate chief physician, Department of Neurosurgery, Tianjin Fourth Central Hospital, Tianjin 300140, China wd5609@ hotmail.com

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610

被引次数

2

摘要：

背景：细胞联合移植、NgR基因沉默及聚乳酸-乙醇酸支架为近年来脊髓神经损伤修复的研究热点。
目的：探讨NgR基因沉默的骨髓间充质干细胞/许旺细胞在聚乳酸-乙醇酸膜上生长及分化的可行性。
方法：将骨髓间充质干细胞、许旺细胞分离、纯化及扩增后，经小分子干扰RNA转染以沉默NgR基因表达，用反转录-聚合酶链反应、Western blot检测两种细胞在转染前后NgR基因/蛋白的表达。在有血清的条件下，按骨髓间充质干细胞组、许旺细胞组及联合组(均以未转染细胞为对照，共6组)分别接种到聚乳酸-乙醇酸膜上，观察各组细胞的贴附、生长、分化情况。
结果与结论：转染小分子干扰RNA后，与转染前相比，实验组NgR基因和蛋白的表达量明显降低(P < 0.05)。经过培养，在聚乳酸-乙醇酸膜上观察与未转染组相比，NgR基因沉默各组均观测到种植细胞的大量贴附和生长。提示NgR基因沉默有促进骨髓间充质干细胞/许旺细胞贴附、生长的作用。

关键词: 聚乳酸-乙醇酸, 骨髓间充质干细胞, 许旺细胞, NgR, 基因沉默, 生物材料

Abstract:

BACKGROUND: Combined cell transplantation, NgR gene silencing and poly-lactic acid-glycolic acid (PLGA) scaffolds have become the research focus of spinal cord injury repair in recent years.
OBJECTIVE: To investigate the feasibility of the growth and differentiation of bone marrow mesenchymal stem cells (BMSCs)/Schwann cells in poly (lactic-co-glycolic acid) (PLGA) membrane after NgR gene silencing.
METHODS: After BMSCs and Schwann cells were isolated, purified and amplified, the two kinds of cells were transfected by small interfering RNA to silence NgR gene expression. The NgR gene/protein expressions in the two kinds of cells before and after transfection were detected by reverse transcription-PCR and Western blot methods. Under serum conditions, the cells were divided into BMSCs group, Schwann cells group and combination group (all untransfected cells as controls, six groups in total). Then, the cells were inoculated onto the PLGA membrane, and their adhesion, growth and differentiation were observed.
RESULTS AND CONCLUSION: The NgR gene and protein expressions of the experimental group were significantly decreased after small interfering RNA transfection (P < 0.05). Compared with the untransfected cells groups after culture, a large number of implanted cells adhered and grew on the PLGA membrane in NgR gene silencing groups. These findings suggest that NgR gene silencing can promote BMSCs/Schwann cells adhesion, growth and differentiation.

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