中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (19): 3538-3541.doi: 10.3969/j.issn.1673-8225.2011.19.028

• 干细胞因子及调控因子 stem cell factors and regulatory factors • 上一篇    下一篇

阿糖胞苷结合神经生长因子体外纯化培养大鼠嗅鞘细胞

刘晓阳，孙建民，崔新刚，蒋振松   

  1. 山东大学附属省立医院脊柱外科，山东省济南市  250021
  • 收稿日期:2010-11-21 修回日期:2011-03-13 出版日期:2011-05-07 发布日期:2011-05-07
  • 通讯作者: 孙建民，博士，主任医师，山东大学附属省立医院脊柱外科，山东省济南市 250021 spine163@163.com
  • 作者简介:刘晓阳★，男，1984年生，山东省青州市人，汉族，在读硕士，主要从事脊髓损伤修复的研究。 shuilanwenqing@163.com
  • 基金资助:

    山东省自然科学基金资助项目(Y2007C010)。

Culture and purification of olfactory ensheathing cells from rats using cytarabine combined with nerve growth factor

Liu Xiao-yang, Sun Jian-min, Cui Xin-gang, Jiang Zhen-song   

  1. Department of Spinal Surgery, Provincial Hospital Affiliated to Shandong University, jinan 250021, Shandong Province, China
  • Received:2010-11-21 Revised:2011-03-13 Online:2011-05-07 Published:2011-05-07
  • Contact: Sun Jian-min, Doctor, Chief physician, Department of Spinal Surgery, Provincial Hospital Affiliated to Shandong University, jinan 250021, Shandong Province, China spine163@163.com
  • About author:Liu Xiao-yang★, Studying for master’s degree, Department of Spinal Surgery, Provincial Hospital Affiliated to Shandong University, jinan 250021, Shandong Province, China shuilanwenqing@163.com
  • Supported by:

    the Natural Science Foundation of Shandong Province, No. Y2007C010*

PDF

287

被引次数

2

摘要：

背景：由于嗅鞘细胞终生具有成髓鞘能力，如何利用简单而又经济的方法获得大量较高纯度的嗅鞘细胞是脊髓损伤研究的热点。
目的：分析阿糖胞苷结合神经生长因子对大鼠嗅球源性嗅鞘细胞纯化培养的可行性。
方法：将差速贴壁后的大鼠嗅鞘细胞首先用含10 mg/L阿糖胞苷的完全培养基培养24 h，然后用含体积分数为1%胎牛血清和25 μg/L神经生长因子的DMEM/F12培养基进行体外培养。观察嗅鞘细胞的生长变化，采用形态学和免疫组化染色对细胞及其纯度进行鉴定。
结果与结论：体外培养的大鼠嗅鞘细胞神经生长因子受体NGFRp75染色呈阳性反应，细胞呈双极和三级，伸出细长突出，并渐结成网状，在体外培养的第7天纯度为95%，第9天时为90%，且形态良好。结果提示阿糖胞苷结合神经生长因子是一种简单实用的嗅鞘细胞纯化培养方法。

关键词: 嗅鞘细胞, 细胞培养, 阿糖胞苷, 神经生长因子, 大鼠

Abstract:

BACKGROUND: Because of the continual capcity of myelination, olfactory ensheathing cells (OECs) are preferred by numerous researchers. The first step is to gain a great quantity of purified OECs simply and economically. The following provides a feasible method to culture OECs.
OBJECTIVE: To explore a simple and pragmatic method to obtain sufficient OECs from neonatal rats by using cytarabine combined with nerve growth factor (NGF).
METHODS: After different attachment, DMEM/F12 culture solution including 10 mg/L cytarabine was used to culture OECs for 48 hours. Then, OECs were cultured in DMEM/F12 solution with 25 μg/L NGF and 1% fetal bovine serum. The conditions and growth degree of OECs were observed and P75 immunocytochemistry was used to estimate the cell purity.
RESULTS AND CONCLUSION: OECs were positive after P75 immunocytochemistry. They appeared to be bipolar or tripolar cells and their processes formed a network in vitro. The purity of OECs in good conditions reached about 95% on 7 days and 90% on 9 days. The method of using cytarabine combined with NGF can culture and purify OECs simply and effectively.

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