中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (40): 7411-7414.doi: 10.3969/j.issn.1673-8225.2010.40.001

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

不同方法诱导成人骨髓间充质干细胞向神经元样细胞分化的比较

张玉红1，韩小改1，辛秀银2   

  1. 1河南省红十字血液中心，河南省郑州市  450012；  2河南省肿瘤医院头颈科室，河南省郑州市  450006
  • 出版日期:2010-10-01 发布日期:2010-10-01
  • 作者简介:张玉红，女，1976年生，河南省郑州市人，汉族，2004年华中科技大学毕业，主管护师，主要从事血液成分制备方面的研究。 zhangyuhong568@163.com

A comparison of various methods for inducing adult bone marrow mesenchymal stem cells differentiation into neuron-like cells

Zhang Yu-hong1, Han Xiao-gai1, Xin Xiu-yin2   

  1. 1 Henan Province Red Cross Blood Center, Zhengzhou  450012, Henan Province, China; 2 Department of Head and Neck, Henan Cancer Hospital, Zhengzhou  450006, Henan Province, China
  • Online:2010-10-01 Published:2010-10-01
  • About author:Zhang Yu-hong, Nurse-in-charge, Henan Province Red Cross Blood Center, Zhengzhou 450012, Henan Province, China zhangyuhong568@163.com

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312

被引次数

4

摘要：

背景：骨髓间充质干细胞可以在体外通过多种诱导剂诱导分化为神经元样细胞，不同诱导剂的细胞分化率、细胞活力以及Nestin染色阳性细胞率不同。
目的：对比多种诱导剂诱导骨髓间充质干细胞分化神经元样细胞结果，探寻一种较好的诱导剂。
方法：取健康成人骨髓于无菌层流室中，加入淋巴细胞分离液离心取白膜，将细胞置于含碱性成纤维细胞生长因子、表皮生长因子、DMEM、体积分数为10%胎牛血清的培养瓶中，37 ℃ 体积分数为5%CO2培养箱中培养，传至第3代分为β-巯基乙醇+二甲基亚砜组，脑源性神经生长因子+维甲酸组，胶质细胞源性神经营养因子+维甲酸组和碱性成纤维细胞生长因子组分别加入诱导剂，观察细胞型态变化，细胞活力测定，免疫组织化学检测神经细胞标志物。
结果与结论：锥虫蓝染色比较各组细胞活力，胶质细胞源性神经营养因子+维甲酸组细胞活力最高(P < 0.05)，β-巯基乙醇+二甲基亚砜组活力最低(P < 0.05)，脑源性神经生长因子+维甲酸组与碱性成纤维细胞生长因子组细胞活力差别无显著性意义(P > 0.05)。神经细胞标志物表达，胶质细胞源性神经营养因子+维甲酸组阳性率高于其余3组(P < 0.05)。结果提示，成人骨髓间充质干细胞可以诱导分化为神经元样细胞，胶质细胞源性神经营养因子+维甲酸组细胞活力高，神经标志物表达阳性率高(P < 0.05)。

关键词: 骨髓间充质干细胞, 诱导, 分化, 胶质细胞源性神经营养因子, 碱性成纤维细胞生长因子

Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) could be induced to differentiate into neuron-like cells through a variety of inducers in vitro. Different inducers of cell differentiation rate, cell viability, as well as the rate of nestin-positive cells staining are different.
OBJECTIVE: To explore a better inducer by comparing results of BMSCs differentiation into neuron-like cells using multiple inducers.
METHODS: The healthy adult bone marrow was taken in the sterile laminar flow room. Lymphocyte separating medium was added for centrifugation, and the albuginea was obtained. Cells were placed in medium containing basic fibroblast growth factor (bFGF), epidermal growth factor, DMEM and 10% fetal bovine serum in 5% CO2 incubator at 37 ℃. The third passage of cells were divided into β-mercaptoethanol + dimethyl sulfoxide group, brain-derived neurotrophic factor (BDNF) + retinoic acid group, glial cell line-derived neurotrophic factor (GDNF) + retinoic acid group and bFGF group. Inducers were added separately. Cell morphology was observed. Cell viability was measured. Nerve cell markers were detected using immunohistochemistry.
RESULTS AND CONCLUSION: Cell viability was compared in each group using trypan blue staining. Cell viability was greatest in the GDNF + retinoic acid group (P < 0.05), but lowest in the β-mercaptoethanol + dimethyl sulfoxide group (P < 0.05). No significant difference in cell viability was determined between the BDNF + retinoic acid group and bFGF group (P > 0.05). Positive rate of nerve cell markers was greater in the GDNF + retinoic acid group compared with β-mercaptoethanol + dimethyl sulfoxide group, BDNF + retinoic acid group and bFGF group (P < 0.05). These suggest that adult BMSCS could differentiate into neuron-like cells. GDNF + retinoic acid was the optimal induction which showed higher living cell energy and positive immunoreactivity to nerve markers (P < 0.05).

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