中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (27): 4975-4978.doi: 10.3969/j.issn.1673-8225.2010.27.008

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

较高温度下骨髓间充质干细胞的复苏率及复苏后分化

柯俊龙，许志恩，李  华，陈静娟   

  1. 广东医学院附属医院神经内科，广东省广州市  524001
  • 出版日期:2010-07-02 发布日期:2010-07-02
  • 通讯作者: 许志恩，主任医师，广东医学院附属医院神经内科，广东省广州市 52400 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

Recovery rate and postresuscitation differentiation of bone marrow mesenchymal stem cells at high temperature  

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

  1. Department of Neurology, Affiliated Hospital of Guangdong Medical College, Guangzhou  524001, Guangdong Province, China
  • Online:2010-07-02 Published:2010-07-02
  • Contact: Xu Zhi-en, Chief physician, Department of Neurology, Affiliated Hospital of Guangdong Medical College, Guangzhou 524001, Guangdong Province, China zjxuzhien@126.com
  • About author:Ke Jun-long★, Master, Attending physician, Department of Neurology, Affiliated Hospital of Guangdong Medical College, Guangzhou 524001, Guangdong Province, China adressor@163.com

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330

被引次数

2

摘要：

背景：目前骨髓间充质干细胞复苏温度多为37 ℃，有研究已证实在精原细胞中可实行较高温度复苏，骨髓间充质干细胞是否能用较高温度复苏尚不清楚。
目的：观察较高复苏温度对骨髓间充质干细胞复苏率和复苏后分化能力的影响。
方法：贴壁培养法分离大鼠骨髓间充质干细胞，传至第5代后将其冻存。然后分别置于37 ℃、50 ℃水浴中复苏，通过锥虫蓝染色法比较两组复苏率。用成纤维细胞生长因子+表皮生长因子+维甲酸诱导50 ℃水浴中复苏的骨髓间充质干细胞向神经样细胞分化，并通过免疫组化染色鉴定。
结果与结论：37 ℃组复苏率(82.40±0.86)%、50℃组复苏率(87.10±1.08)%，两组复苏率差异有显著性意义(P < 0.01)，50 ℃复苏组细胞加入诱导剂后出现典型神经元样改变，Nestin免疫组化染色阳性。结果提示50 ℃复苏骨髓间充质干细胞较    37 ℃获得较高的复苏率，且不影响其分化能力。

关键词: 温度, 复苏, 50 ℃, 骨髓间充质干细胞, 分化能力

Abstract:

BACKGROUND: At present, temperature of recovery of bone marrow mesenchymal stem cells (BMSCs) is 37 ℃. Previous reports have confirmed that spermatogonia were recovered at a high temperature, but it remains poor understood whether BMSCs were recovered at a high temperature.
OBJECTIVE: To observe the effects of high temperature on recovery rate and differentiation ability of BMSCs.
METHODS: Rat BMSCs were isolated and cultured by adherence method. The fifth passage of BMSCs were cryopreserved, and then recovered at 37 ℃ and 50 ℃ in water bath. The recovery rate was compared using trypan blue staining in both groups. BMSCs were induced to differentiate into neuron-like cells by fibroblast growth factor + epidermal growth factor + ratinoic acid at 50 ℃ in water bath, and identified by immunohistochemical staining.
RESULTS AND CONCLUSION: Recovery rates were respectively (82.40±0.86)% and (87.10±1.08)% in the 37 ℃ and 50 ℃ groups (P < 0.01). In the 50 ℃ group, cells presented neuron-like changes following inducer intervention, and were positive for Nestin in immunohistochemical staining. Results have suggested that the recovery rate in BMSCs was higher in 50 ℃ group compared with 37 ℃ group. Moreover, the differentiation ability was not affected.

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