Kong Xianmin, Tian Shanshan, Chen Tao, Huang Yinghui
School of Biomedical Engineering 【?】, Beijing University of Technology, Beijing, 100124, P.R.China.
Institute of Laser Engineering, Beijing University of Technology, Beijing, 100124, P.R.China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2019 Feb 15;33(2):239-242. doi: 10.7507/1002-1892.201809094.
To preliminary study on the feasibility of constructing three-dimensional (3D) hippocampal neural network by using microfluidic technology.
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.
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 . 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.
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 .
初步研究利用微流控技术构建三维(3D)海马神经网络的可行性。
通过标准湿法蚀刻工艺设计并制作了一种网络图案化微流控芯片。分离并培养新生Sprague Dawley大鼠的原代海马神经元,然后接种到微流控芯片上进行培养。采用免疫荧光染色观察培养3、5和7天时海马神经元的生长情况,并在培养7天时对海马神经元网络进行电生理检测。
结果显示,随着培养时间的延长,海马神经元数量逐渐增加,神经元的神经突也相应增加,并在微流控芯片通道中均匀且规则地分布,表明成功构建了3D海马神经元网络。在培养7天时可检测到海马神经元网络的单通道和多通道自发放电信号,表明神经元网络具有初步的生物学功能。
图案化微流控芯片可使海马神经元沿有限路径生长并形成具有信号转导等相应生物学功能的3D神经元网络,为进一步探索神经元网络的功能奠定了基础。