《中国康复理论与实践》

《中国康复理论与实践》 ›› 2015, Vol. 21 ›› Issue (10): 1145-1150.

• 基础研究 • 上一篇    下一篇

不同胎龄胎鼠脊髓源性神经干细胞特性比较

南伟 1a,2，冯东亮 1a,2，周开升 1a,2，朱彦东 1a,2，汪静 1a,2，田婷 1b，江龙 3，龙在云 3，伍亚民 3，张海鸿 1a   

  1. 1.兰州大学第二医院，a.骨科；b.神经内科，甘肃兰州市 730030；2.甘肃省骨关节疾病研究重点实验室，甘肃兰州市 730030；3.第三军医大学大坪医院野战外科研究所三室，创伤烧伤与复合伤国家重点实验室，重庆市 400042。
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2015-10-25 发布日期:2015-10-25

Differentiation of Spinal Neural Stem Cells from Various Gestational Aged Fetal Rats

NAN Wei1a,2, FENG Dong- liang1a,2, ZHOU Kai- sheng1a,2, ZHU Yan- dong1a,2, WANG Jing1a,2, TIAN Ting1b, JIANG Long3, LONG Zai-yun3, WU Ya-min3, ZHANG Hai-hong1a   

  1. 1. a. Department of Orthopedics; b. Department of Neurology, the Second Clinical Medical College of Lanzhou University, Lanzhou, Gansu 730030, China; 2. Key Laboratory of Orthopedics of Gansu Province, Lanzhou, Gansu 730030, China; 3. State Key Laboratory of Trauma, Burns and Combined Injury, the third Department of Research Institute of Surgery, Daping Hospital, Third Military Medical University, Chongqing 400042, China
  • Received:1900-01-01 Revised:1900-01-01 Published:2015-10-25 Online:2015-10-25

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摘要： 目的 探讨不同胎龄胎鼠脊髓源性神经干细胞(NSCs)的增殖分化能力。方法 Sprague-Dawley孕鼠分为 A组(孕 12d)、B组(孕14 d)、C组(孕16 d)，酶消化法结合机械分离法提取细胞，检测不同时间各组神经干细胞球的直径、数量，CCK-8法绘制细胞生长曲线。BrdU/Nestin免疫荧光染色鉴定NSCs。10%胎牛血清诱导分化后行β-tubulinⅢ、胶质纤维酸性蛋白(GFAP)免疫荧光染色，比较各组神经元的分化比例。结果 各组细胞 BrdU、Nestin、β-tubulinⅢ、GFAP免疫荧光染色均为阳性。B组β-tubulinⅢ染色阳性的比例最高；B、C两组各时间点神经球直径、数量及神经元的分化比例高于 A组(P<0.05)。第 3代 NSCs培养第 5天，B组细胞吸光度高于C组(P<0.05)。结论 孕14 d左右取材培养的脊髓神经干细胞生物学特性稳定，增殖活力强，诱导分化后神经元的分化比例较高。

关键词: 神经干细胞, 脊髓, 神经元, 星形胶质细胞, 原浆型星形胶质细胞, 大鼠

Abstract: Objective To study the differentiation and proliferation ability of the spinal neural stem cells (NSCs) at different gestational ages in fetal rats. Methods Sprague-Dawley fetal rats were divided into group A (12 days of pregnancy), group B (14 days of pregnancy) and group C (16 days of pregnancy). NSCs were separated with enzyme-assisted microdissection. The diameter and numbers of NSCs balls were measured at different time. The cell growth curve was drawn with CCK8 colorimeter. NSCs were identified with BrdU/Nestin immunohistochemical staining. They were induced with 10% fetal bovine serum for 10 days, and the expression of β-tubulinⅢ and glial fibrillary acidic protein was detected with immunocytochemistry. Results There were cells expressed BrdU, Nestin, β-tubulinⅢ and GFAP in all the group. The most cells (22.74±0.79%) expressed β-tubulinⅢ in the group B, but no significant difference between group B and group C. The cell vitality on the 5th day of third-generation neural stem cells was the most in group B. Conclusion For enzyme-assisted microdissection, it may obtain more neurons to isolate the neural stem cells from 14 days of pregnancy pregnant rats.

Key words: neural stem cells, spinal cord, neurons, astrocyte, protoplasmic astrocyte, rats