中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (14): 2481-2484.doi: 10.3969/j.issn.1673-8225.2010.14.003　

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

大鼠骨髓间充质干细胞体外向神经元样细胞的诱导

谷艳娇, 李  博, 高志安   

  1. 辽宁医学院病理学教研室，辽宁省锦州市  121001
  • 出版日期:2010-04-02 发布日期:2010-04-02
  • 通讯作者: 高志安，博士，教授，辽宁医学院病理学教研室，辽宁省锦州市 121001
  • 作者简介:谷艳娇★，女，1970年生，辽宁省本溪市人，汉族，2009年辽宁医学院毕业，硕士，主治医师，主从事诊断病理学。 LHD373@sohu.com

Induction and differentiation of rat bone marrow mesenchymal stem cells into neuron-like cells in vitro

Gu Yan-jiao, Li Bo, Gao Zhi-an   

  1. Department of Pathology, Liaoning Medical University, Jinzhou  121001，Liaoning Province, China
  • Online:2010-04-02 Published:2010-04-02
  • Contact: Gao Zhi-an, Doctor, Professor, Department of Pathology, Liaoning Medical University, Jinzhou 121001，Liaoning Province, China
  • About author:Gu Yan-jiao, Master, Attending physician, Department of Pathology, Liaoning Medical University, Jinzhou 121001，Liaoning Province, China LHD373@sohu.com

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368

被引次数

22

摘要：

背景：选择合适的诱导方法诱导骨髓间充质干细胞为神经元样细胞是临床治疗神经损伤的关键。
目的：在大鼠骨髓间充质干细胞中加入大鼠齿状回蛋白提取物，观察其诱导分化为神经元样细胞的可行性。
方法：应用20，40，60，80 mg/L齿状回蛋白提取物诱导分化大鼠骨髓间充质干细胞，蛋白印记检测神经元分化情况，筛选最佳剂量。收集诱导后的神经元样细胞，倒置显微镜下观察分化后细胞的形态学变化，用蛋白印记技术检测NSE、NeuN的表达，免疫荧光技术检测细胞内NeuN的表达。
结果与结论：诱导分化7 d后，经蛋白印记检测可见60 mg/L的齿状回蛋白提取物为最佳诱导剂量，随质量浓度的增加NSE与NeuN的表达量逐渐增加，当质量浓度为80 mg/L时，细胞表达的NSE与NeuN减少；选择60 mg/L齿状回蛋白提取物诱导分化骨髓间充质干细胞，诱导后的细胞变长，有突触；免疫荧光NeuN染色发现，诱导分化后的神经元样细胞有阳性表达。结果提示齿状回提取物是一种可以诱导骨髓间充质干细胞分化的有效生物诱导剂。

关键词: 齿状回提取物, 骨髓间充质干细胞, 诱导分化, 神经元样细胞, 生物诱导剂

Abstract:

BACKGROUND: It is a key to choose an appropriate method to trans-differentiated bone marrow mesenchymal stem cells (BMSCs) into neuron-like cells for clinical therapy of neural system injury.
OBJECTIVE: To observe the feasibility of the differentiation of rat BMSCs supplemented with rat dentate gyrus extract (DGE) into neuron-like cells.
METHODS: DGE was applied to induce rat BMSC trans-differentiation, using 20, 40, 60, 80 mg/L protein concentration. Neuron differentiation was measured using Western blot to screen an optimal dosage. After trans-differentiation, different concentration treated cells were collected. Morphology change was observed following differentiation under an inverted microscope. Neuron specific enolase (NSE) and NeuN expression was determined using Western blot. NeuN expression in cells was detected using immunofluorescence technique.
RESULTS AND CONCLUSION: After 7 days, 60 mg/L DGE was the optimal induction dose detected by Western blot. With increased concentration, NSE and NeuN expression was increased. At 80 mg/L mass concentration, NSE and NeuN expression was reduced. At 60 mg/L DGE, BMSCs following induction became long, with synapses. Immunofluorescence NeuN staining found that neuron-like cells were positive for NeuN following induction. Results indicated that DGE is an effective biological inductor that can induce BMSC differentiation.

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