中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (14): 2617-2620.doi: 10.3969/j.issn.1673-8225.2010.14.030

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

新生C57小鼠耳蜗螺旋神经节细胞的原代培养

王庭阔，孙  虹   

  1. 中南大学湘雅三医院耳鼻咽喉-头颈外科，湖南省长沙市  410013
  • 出版日期:2010-04-02 发布日期:2010-04-02
  • 通讯作者: 孙 虹，教授，中南大学湘雅三医院耳鼻咽喉-头颈外科，湖南省长沙市 410013 shjhaj@vip163.com
  • 作者简介:王庭阔☆，男，1971年生，湖南省沅陵县人，2007级中南大学湘雅三医院在读博士，主治医师，主要从事耳科临床的研究。 wtk711112@163.com
  • 基金资助:

    国家自然科学基金(30371531)，耳蜗兴奋性损伤后神经营养素-3对神经元的保护作用研究；湖南省自然科学基金(02JJY2050)，神经营养素-3和尼莫地平对抗耳蜗兴奋性损害的实验研究；湖南省卫生厅科研基金(B2006-063)，羟基磷灰石纳米载体介导神经营养素3基因治疗神经性耳聋；湖南省科技厅科技计划重点项目(2007SK2001)，羟基磷灰石纳米载体介导内耳基因转染的关键技术研究；国家自然科学基金(30772402)，羟基磷灰石纳米粒经完整圆窗膜向内耳转染NT-3基因的研究。

Primary culture of spiral ganglion cells from cochlea of neonatal C57 mice 

Wang Ting-kuo, Sun Hong   

  1. Department of Otorhinolaryngology-Head ＆ Neck Surgery, Third Xiangya Hospital, Central South University, Changsha   410013, Hunan Province, China
  • Online:2010-04-02 Published:2010-04-02
  • Contact: Sun Hong, Professor, Department of Otorhinolaryngology-Head ＆ Neck Surgery, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan Province, China shjhaj@vip163.com
  • About author:Wang Ting-kuo☆, Studying for doctorate, Attending physician, Department of Otorhinolaryngology-Head ＆ Neck Surgery, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan Province, China wtk711112@163.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30371531*，30772402*; the Natural Science Foundation of Hunan Province, No. 02JJY2050*; the Scientific Research Foundation of Health Department of Hunan Province, No. B2006-063*; the Key Program of Science and Technology Project of Department of Science and Technology of Hunan Province, No. 2007SK2001*

PDF

473

被引次数

2

摘要：

背景：目前在原代细胞培养领域内，对耳蜗螺旋神经节细胞培养条件的报道各有差异，个别方法重复性较差，不利于实际应用。
目的：原代培养并鉴定新生C57小鼠螺旋神经节细胞。
方法：显微解剖分离新生C57小鼠蜗轴组织，经胰酶消化+差速贴壁+化学药物相结合方法培养；倒置相差显微镜及苏木精-伊红染色观察细胞生长状态，免疫组织化学染色鉴别细胞来源。
结果与结论：蜗轴组织细胞纯化后，胞体呈椭圆形或三角形，有细长的突起，Nuen染色胞核呈棕黄色阳性反应，β3-Tubulin染色细胞胞浆与轴突均呈棕黄色阳性反应。提示实验成功培养出小鼠螺旋神经节细胞。

关键词: 螺旋神经节细胞, 细胞模型, 原代培养, 小鼠, 耳蜗

Abstract:

BACKGROUND: At present, there are differences in reporting culture condition of cochleal spiral ganglion cells (SGCs) in circle of primary cell culture. Individual method has poor repeatability, and is not beneficial for practical application.
OBJECTIVE: To primary culture and evaluate SGCs of neonatal C57 mice.
METHODS: Tissues from cochlear modiolus were acquired from neonatal C57 mice by microanatomy. SGCs from cochlear modiolus were cultured by trypsin digestion, differential velocity adherent technique combined with chemicals. Cell growth was observed by inverted phase contrast microscope and hematoxylin and eosin staining. Immunocytochemistry was employed to classify cell types. 
RESULTS AND CONCLUSION: After purification, the cell body of SGCs from cochlear modiolus was ellipse or triangle, with slender processes in cytoplasm. Nuclei were positive for Nuen (stained brown). Cytoplasm and axon were positive for β3-Tubulin (stained brown). These indicated that mouse SGCs were successfully cultured.

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