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作者:

Kong, X. (Kong, X..) | Tian, S. (Tian, S..) | Chen, T. (Chen, T..) | Huang, Y. (Huang, Y..) (学者:黄艳)

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摘要:

目的: 初步探讨利用微流控技术体外构建三维海马神经元网络的可行性。. 方法: 设计并采用标准湿法腐蚀工艺制备网络图案化的微流控芯片。取新生 SD 大鼠丘脑组织,分离培养原代海马神经元后,接种于微流控芯片上进行培养。3、5、7 d 时行免疫荧光染色,观察海马神经元生长情况;7 d 时行海马神经元网络电生理检测。. 结果: 荧光显微镜下观察显示,随培养时间延长,海马神经元数量逐渐增加,且神经元突起亦相应增多、增长,并且均匀规则分布在微流控芯片通道,提示成功构建三维海马神经元网络。培养 7 d 时可以检测到海马神经元网络的单通道及多通道自发放电信号,提示神经元网络具有初步的生物学功能。. 结论: 图案化微流控芯片可以使海马神经元按照局限路径生长,从而形成三维神经元网络,并且具有信号传导等相应的生物学功能。.Objective: To preliminary study on the feasibility of constructing three-dimensional (3D) hippocampal neural network in vitro by using microfluidic technology. Methods: A network patterned microfluidic chip was designed and fabricated by standard wet etching process. The primary hippocampal neurons of neonatal Sprague Dawley rats were isolated and cultured, and then inoculated on microfluidic chip for culture. Immunofluorescence staining was used to observe the growth of hippocampal neurons at 3, 5, and 7 days of culture and electrophysiological detection of hippocampal neuron network at 7 days of culture. Results: The results showed that the number of hippocampal neurons increased gradually with the prolongation of culture time, and the neurite of neurons increased accordingly, and distributed uniformly and regularly in microfluidic chip channels, suggesting that the 3D hippocampal neuron network was successfully constructed in vitro. Single and multi-channel spontaneous firing signals of hippocampal neuronal networks could be detected at 7 days of culture, suggesting that neuronal networks had preliminary biological functions. Conclusion: Patterned microfluidic chips can make hippocampal neurons grow along limited paths and form 3D neuron networks with corresponding biological functions such as signal transduction, which lays a foundation for further exploring the function of neuron networks in vitro.

关键词:

hippocampal neuron; Microfluidic chip; neuron network; three-dimensional culture

作者机构:

  • [ 1 ] [Kong, X.]School of Biomedical Engineering 【?】, Beijing University of Technology, Beijing, 100124, China
  • [ 2 ] [Kong, X.]School of Biomedical Engineering 【?】, Beijing University of Technology, Beijing, 100124, China
  • [ 3 ] [Tian, S.]Institute of Laser Engineering, Beijing University of Technology, Beijing, 100124, China
  • [ 4 ] [Chen, T.]Institute of Laser Engineering, Beijing University of Technology, Beijing, 100124, China
  • [ 5 ] [Huang, Y.]School of Biomedical Engineering 【?】, Beijing University of Technology, Beijing, 100124, China
  • [ 6 ] [Huang, Y.]School of Biomedical Engineering 【?】, Beijing University of Technology, Beijing, 100124, China

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来源 :

Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = Chinese journal of reparative and reconstructive surgery

ISSN: 1002-1892

年份: 2019

期: 2

卷: 33

页码: 239-242

被引次数:

WoS核心集被引频次: 0

SCOPUS被引频次: 1

ESI高被引论文在榜: 0 展开所有

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