口腔医学研究

口腔医学研究 ›› 2018, Vol. 34 ›› Issue (4): 411-413.DOI: 10.13701/j.cnki.kqyxyj.2018.04.018

• 口腔颌面外科学研究 • 上一篇    下一篇

新生SD大鼠三叉神经元细胞的体外原代培养及鉴定

邓超, 张鹤, 薛进朗, 王丽婵, 郭家奕, 程慧欣, 陈传俊*   

  1. 皖南医学院口腔医学院 安徽 芜湖 241000
  • 收稿日期:2017-10-16 出版日期:2018-04-28 发布日期:2018-04-25
  • 通讯作者: 陈传俊,E-mail:ccj6318@sina.com
  • 作者简介:邓超(1984~ ),男,安徽南陵人,讲师,博士,主要从事组织再生研究工作。
  • 基金资助:
    国家自然科学基金(编号:81550037)安徽省教育厅高校自然科学研究项目(编号:KJ2017A255)皖南医学院博士科研启动基金(编号:WK2014RC03)皖南医学院大学生科研基金(编号:WK2016S10)

Primary Culture and Identification of Trigeminal Neurons in Neonatal SD Rats.

DENG Chao, ZHANG He, XUE Jin-lang, WANG Li-chan, GUO Jia-yi, CHENG Hui-xin, CHEN Chuan-jun*   

  1. School of Stomatology, Wannan Medical College, Wuhu 241000, China.
  • Received:2017-10-16 Online:2018-04-28 Published:2018-04-25

PDF (PC)

401

被引次数

3

摘要： 目的:建立新生SD大鼠三叉神经元细胞体外原代培养模型,为三叉神经的细胞学研究提供实验基础。方法:选取新生24 h以内SD大鼠24只,剥离脑组织,手术显微镜下分离大鼠三叉神经节,无菌剪碎后采用胰蛋白酶与DNA酶混合分次消化和机械吹打相结合的方法制备神经细胞悬液,接种于6孔板,倒置相差显微镜下观察细胞状态;免疫荧光染色鉴定神经元特异性抗原NeuN、β-tubulin Ⅲ的表达。结果:分离出来的三叉神经细胞4 h后贴壁生长,并长出突起,随时间增加,突起变长并交错呈网络,培养至3~7 d神经细胞状态最佳,随后逐渐凋亡,体外培养的神经细胞可生长15d,无增殖传代的能力;神经元特异性抗原NeuN、β-tubulin Ⅲ的表达阳性。结论:胰蛋白酶与DNA酶混合分次消化法可分离出大量三叉神经元细胞,为后续的细胞学研究提供实验基础。

关键词: 三叉神经元, 细胞培养, 免疫荧光

Abstract: Objective: To establish a primary culture model of trigeminal neurons in newborn SD rats and provide experimental basis for the study of trigeminal nerve cytology. Methods: Twenty-four new born SD rats were selected and the trigeminal ganglion was isolated under the microscope. Neural cell suspension was prepared by trypsin and DNase digestion and mechanical blowing. The expression of neuron specific antigen NeuN and β-tubulinⅢ was identified by immunofluorescence staining. Results: Nerve cells were adherent growth, and reached protrusions. With time, the protrusions became longer and staggered network. The cells reached the best status after cultured for 3-7 days, then gradually apoptosis. The cells could culture for 15 days without ability to pass on, and the expressions of neuronal specific antigen NeuN and β-tubulinⅢ were positive. Conclusion: The combination of trypsin and DNase digestion can isolate a large number of trigeminal neurons, which provides the experimental basis for cytology research.

Key words: Trigeminal neurons, Cell culture, Immunofluorescence