中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (49): 9174-9177.doi: 10.3969/j.issn.1673-8225.2011.49.012

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

促红细胞生成素体外培养鼠胚脑皮质神经干细胞的分化

袁丽丽1，杜红梅2，管英俊2，孔佑华1   

  1. 1济宁医学院组织胚胎学教研室，山东省济宁市   272067
    2潍坊医学院组织胚胎学教研室，山东省潍坊市  261053
  • 收稿日期:2011-06-18 修回日期:2011-10-20 出版日期:2011-12-03 发布日期:2011-12-03
  • 作者简介:袁丽丽★，女，1979年生，山东省青岛市人，汉族，2007年潍坊医学院毕业，硕士，讲师，主要从事神经再生与修复研究。 liliyuan79@yahoo.com.cn
  • 基金资助:

    济宁医学院青年科研基金项目资助项目(2008jnqn-25)，课题名称：EPO对体外鼠胚脑皮质神经干细胞增殖、分化及凋亡的影响。

Differentiation of rat embryonic cerebral cortex neural stem cells cultured by erythropoietin in vitro

Yuan Li-li1, Du Hong-mei2, Guan Ying-jun2, Kong You-hua1   

  1. 1Department of Histology and Embryology of Jining Medical College, Jining  272067, Shandong Province, China
    2Department of Histology and Embryology of Weifang Medical College, Weifang  261053, Shandong Province, China
  • Received:2011-06-18 Revised:2011-10-20 Online:2011-12-03 Published:2011-12-03
  • About author:Yuan Li-li★, Master, Lecturer, Department of Histology and Embryology of Jining Medical College, Jining 272067, Shandong Province, China liliyuan79@yahoo.com.cn
  • Supported by:

    Youth Research Foundation of Jining Medical University, No. 2008jnqn-25*

PDF

512

被引次数

16

摘要：

背景：促红细胞生成素最早作为生长因子被认识，近些年来其对中枢神经系统保护作用的体内研究较多。
目的：观察促红细胞生成素对体外培养大鼠胚脑皮质神经干细胞凋亡及分化的影响。
方法：无菌条件下取孕14 d SD大鼠胚脑皮质，体外先悬浮增殖培养后贴壁诱导分化。巢蛋白免疫细胞荧光染色检测神经干细胞，以微管相关蛋白2、神经胶质原纤维酸性蛋白检测神经干细胞分化。取第3代神经干细胞向培养基中添加0.5，5，50，500 U/mL的促红细胞生成素，另设不添加促红细胞生成素的对照组。应用半胱氨酸天冬氨酸蛋白酶3检测神经干细胞凋亡，微管相关蛋白2检测神经干细胞向神经元方向的分化。
结果与结论：加入≥5 U/mL促红细胞生成素，神经球内半胱氨酸天冬氨酸蛋白酶3表达显著下降(P < 0.01)，分化培养后微管相关蛋白2阳性细胞较对照组明显升高(P < 0.01)。提示促红细胞生成素可降低体外培养鼠胚脑皮质神经干细胞的凋亡率，促进其神经干细胞向神经元方向分化。

关键词: 促红细胞生成素, 神经干细胞, 培养, 凋亡, 分化

Abstract:

BACKGROUND: Erythropoietin (FPO) is early known as a kind of growth factor. In recent years, there are many studies about the protective effect of erythropoietin on the central nervous system in vivo.
OBJECTIVE: To explore the effects of EPO on the apoptosis and differentiation of rat embryonic cerebral cortex neural stem cells in vitro.
METHODS: The embryonic cerebral cortex were isolate from Sprague-Dawley rats that pregnant for 14 days under sterile conditions, the cells were cultured and differentiated in the suspension medium and then adherent induction. The neural stem cells were detected by nestin immune cell fluorescence staining, microtubule-associated protein 2 (MAP-2) and glia fibrillary acid protein was used to detect the differentiation of neural stem cells. The third passage of neural stem cells were obtained and 0.5, 5, 50, 500 U/mL EPO were added into the medium, and in the control group, there was no EPO adding. Caspase-3 was used to detect the apoptosis of neural stem cells, and MAP-2 was used to detect the differentiation of neural stem cells to neuron.
RESULTS AND CONCLUSION: The expression of the caspase-3 in neural sphere was decreased after added EPO (≥5 U/mL), and the expression of the MAP-2 positive cells was increased obviously after differentiation (P < 0.01). The present results suggest that EPO can decrease the apoptosis of the neural stem cells and promote neural stem cells to differentiate into neurons in vitro.

中图分类号: