中国组织工程研究

中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (45): 7266-7272.doi: 10.3969/j.issn.2095-4344.2014.45.010

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

新生小鼠海马、嗅球及皮质神经干细胞的分离培养及鉴定

马浚宁1，2，高俊玮1，侯博儒1，任海军1，2，陈四化1，刘吉星1，严贵忠1   

  1. 1兰州大学第二临床医院神经外科，甘肃省兰州市 730000； 2兰州大学生命科学学院，甘肃省兰州市 730000
  • 出版日期:2014-11-05 发布日期:2014-11-05
  • 通讯作者: 任海军，教授，硕士生导师，主任医师，兰州大学第二临床医院神经外科，甘肃省兰州市 730000
  • 作者简介:马浚宁，男，1990年生，青海省西宁市人，回族，兰州大学第二临床医学院在读硕士，主要从事神经干细胞移植治疗神经系统疾病的研究。
  • 基金资助:

    甘肃省自然科学基金(1208RJZA184)

Isolation and identification of neural stem cells from newborn mouse hippocampus, olfactory bulb and cortex

Ma Jun-ning1, 2, Gao Jun-wei1, Hou Bo-ru1, Ren Hai-jun1, 2, Chen Si-hua1, Liu Ji-xing1, Yan Gui-zhong1   

  1. 1Department of Neurosurgery, Lanzhou University Second Hospital, Lanzhou 730000, Gansu Province, China; 2School of Life Sciences, Lanzhou University, Lanzhou 730000, Gansu Province, China
  • Online:2014-11-05 Published:2014-11-05
  • Contact: Ren Hai-jun, Professor, Master’s supervisor, Chief physician, Department of Neurosurgery, Lanzhou University Second Hospital, Lanzhou 730000, Gansu Province, China
  • About author:Ma Jun-ning, Studying for master’s degree, Department of Neurosurgery, Lanzhou University Second Hospital, Lanzhou 730000, Gansu Province, China; School of Life Sciences, Lanzhou University, Lanzhou 730000, Gansu Province, China
  • Supported by:

     the Natural Science Foundation of Gansu Province, No. 1208RJZA184

PDF

680

被引次数

16

摘要：

背景：从体外分离培养出高纯度、生物学性能均一的神经干细胞，建立起一套完整的神经干细胞培养体系，是进行神经干细胞研究的基础。

 

目的：建立新生小鼠海马、嗅球、皮质组织神经干细胞的分离培养体系，并对其生物学特性进行分析。

 

方法：分离新生昆明小鼠海马、嗅球、皮质组织，采用机械分离和胰酶消化法提取原代神经干细胞。采用无血清培养技术、机械吹打和酶消化法进行传代培养神经干细胞。以体积分数为10%的胎牛血清诱导分化神经干细胞。对神经干细胞及其分化产物行CD133、巢蛋白、β-微管蛋白Ⅲ、胶质纤维酸性蛋白免疫荧光染色鉴定。

 

结果与结论：从新生小鼠海马、嗅球、皮质可提取出具有自我更新和多向分化能力的神经干细胞，经巢蛋白、CD133免疫荧光染色检测呈阳性；神经干细胞经胎牛血清诱导后可分化为β-微管蛋白Ⅲ、胶质纤维酸性蛋白阳性细胞，并证实染色阳性细胞为神经元和星形胶质细胞。该实验建立了一套神经干细胞体外分离培养、纯化、鉴定、诱导分化方案，为后续神经干细胞研究的顺利进行奠定了实验基础。

 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程


全文链接：

关键词: 干细胞, 培养, 神经干细胞, 神经球, 海马, 嗅球, 皮质, CD133, 巢蛋白, 神经干细胞库, 甘肃省自然科学基金

Abstract:

BACKGROUND: To in vitro isolate neural stem cells with high purity and uniform biological properties and to establish a complete set of neural stem cell culture system is the basis for neural stem cell research.

OBJECTIVE: To establish an isolation and culture system for neural stem cells from newborn mouse hippocampus, olfactory bulb and cortex and to analyze the biological properties of cells.
METHODS: Neural stem cells were isolated from the hippocampus, olfactory bulb and cortex tissue of newborn Kunming mice by mechanical separation and trypsin digestion. Serum-free culture technology, mechanical pipetting and trypsin digestion were used for subculture of neural stem cells. 10% fetal bovine serum was used to induce differentiation of neural stem cells. Neural stem cells and their differentiated products were identified by immunofluorescent staining of Nestin, CD133, β-TubulinIII, glial fibrillary acidic protein.
RESULTS AND CONCLUSION: The neural stem cell obtained from newborn mouse hippocampus, olfactory bulb and cortex had the capacity of self-renewal and differentiation which were positive for Nestin and CD133. After induction with fetal bovine serum, neural stem cell could differentiation to β-tubulinIII or glial fibrillary acidic protein positive cells that were neurons and astrocytes. This experiment has successfully established the neural stem cell isolation, culture, identification and induction system, providing experimental basis for subsequent studies of neural stem cells.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程


全文链接：

Key words: neural stem cells, mice, hippocampus, olfactory bulb

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