中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (32): 5908-5912.doi: 10.3969/j.issn.1673-8225.2010.32.004

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

细胞密度与骨髓间充质干细胞向神经元样细胞的分化

柯俊龙，许志恩，李  华，陈静娟，梁  维   

  1. 广东医学院附属医院神经内科, 广东省湛江市  524001
  • 出版日期:2010-08-06 发布日期:2010-08-06
  • 通讯作者: 许志恩，主任医师，广东医学院附属医院神经内科，广东省湛江市 524001 zjxuzhien@126.com
  • 作者简介:柯俊龙★，男，1977年生，广东省化州市人，汉族，2010年广东医学院毕业，硕士，主治医师，主要从事干细胞研究和脑血管疾病研究。 adressor@163. com

Effect of cell density on differentiation of bone marrow mesenchymal stem cells into neuronal-like cells 

Ke Jun-long, Xu Zhi-en, Li Hua, Chen Jing-juan, Liang Wei   

  1. Department of Neurology, Affiliated Hospital of Guangdong Medical College，Zhanjiang   524001, Guangdong Province, China
  • Online:2010-08-06 Published:2010-08-06
  • Contact: Xu Zhi-en, Chief physician, Department of Neurology, Affiliated Hospital of Guangdong Medical College, Zhanjiang 524001, Guangdong Province, China zjxuzhien @126．com
  • About author:Ke Jun-long★, Master, Attending physician, Department of Neurology, Affiliated Hospital of Guangdong Medical College, Zhanjiang 524001, Guangdong Province, China adressor@163.com

PDF

454

被引次数

8

摘要：

背景：细胞种植密度是影响干细胞分化的因素之一，对于细胞种植密度在骨髓间充质干细胞向神经元样细胞分化过程中的作用尚缺乏深入研究。
目的：观察细胞种植密度对骨髓间充质干细胞诱导向神经元样细胞分化的影响。
方法：采用贴壁培养法分离大鼠骨髓间充质干细胞，传至第4代后将其按2×102，2×103，4×103，8×103，2×104，4×104/cm2种植于六孔板，每组均加入碱性成纤维细胞生长因子+表皮生长因子+维甲酸诱导向神经元样细胞分化，并通过免疫组织化学染色鉴定，计算每组细胞出现神经元样细胞的比例，比较各组的分化率。
结果与结论：各组骨髓间充质干细胞加入诱导剂后均出现神经元样细胞，Nestin、NSE、GFAP细胞化学染色呈阳性。不同种植密度组出现神经元样细胞比例不同，以8×103/cm2组神经元样细胞比例最高，且神经元样存活时间最长，达7 d。结果说明骨髓间充质干细胞向神经元样细胞分化与细胞接种密度有关，过高或过低细胞密度均不利分化。

关键词: 骨髓间充质干细胞, 细胞密度, 分化, 神经元样细胞, 干细胞培养与分化

Abstract:

BACKGROUND: Cell plant density is one of the factors that affect stem cell differentiation. To date, there is still lack of further study about the effect of cell plant density on bone marrow mesenchymal stem cells differentiation into neuronal-like cells.
OBJECTIVE: To investigate the effect of cell density on the differentiation of rat bone marrow mesenchymal stem cells into neuronal-like cells.
METHODS: Rat bone marrow mesenchymal stem cells were isolated by use of adherent culture method. Following four passages, cells were cultured in six-well plates at a density of 2×102/cm2, 2×103/cm2, and 4×103/cm2, 8×103/cm2, 2×104/cm2, 4×104/cm2. Cells in each group were differentiated into neuronal-like cells under the induction of basic fibroblast growth factor + epidermal growth factor + RA, and were identified by immunohistochemical staining method. The proportion of neuronal-like cells in each group was calculated, and the differentiation rate was compared.
RESULTS AND CONCLUSION: Neuronal-like cells were observed after adding inducer in bone marrow mesenchymal stem cells, and Nestin, neuron-specific enolase, glial fibrillary acidic protein staining was positive. Different cell plant densities induced different ratios of neuron-like cells. 8×103/cm2 plant density yielded highest proportion of neuronal-like cells and can survive as long as 7 days. The results demonstrated that the differentiation of bone mesenchymal stem cells into neuron-like cells was correlated with cell plant density. Too high or too low cell plant density was detrimental to differentiation.

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