Recording axonal conduction to evaluate the integration of pluripotent cell-derived neurons into a neuronal network
Recording axonal conduction to evaluate the integration of pluripotent cell-derived neurons into a neuronal network
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记录轴突传导以评估多能细胞源性神经元整合到神经元网络中
DOI：
10.1007/s10544-015-9997-y
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发表时间：
2015
影响因子：
2.8
通讯作者：
Yasuhiko Jimbo
中科院分区：
文献类型：
--
作者：
Kenta Shimba;Koji Sakai;Yuzo Takayama;Kiyoshi Kotani;Yasuhiko Jimbo
干细胞移植是治疗神经退行性疾病的一种很有前途的方法，许多体外模型已被开发用于研究移植神经元和宿主神经元网络之间的相互作用，以促进药物发现。然而，缺乏能够评估干细胞整合到宿主神经元网络中的过程的方法。在这项研究中，我们应用了轴突传导为基础的分析，原代和分化的神经元的共培养研究。将小鼠皮层神经元和从P19胚胎癌细胞分化的神经元细胞（多能干细胞的早期神经分化的模型）在微制造装置中共培养。这些细胞的胞体被共培养装置分离，但它们的轴突能够通过微通道伸长，然后形成突触接触。通过嵌入微通道底部的微电极记录这些轴突的动作电位，并将其分类为代表单个轴突的簇。皮层神经元轴突数目在体外培养14天后逐渐增加，14天后逐渐减少，而P19神经元轴突数目在整个培养过程中逐渐增加。网络爆发分析表明，P19神经元在体外14天后参与了大约80%的爆发活动。有趣的是，P19神经元的轴突传导延迟明显大于皮质神经元，这表明它们的轴突存在一些生理差异。这些结果表明，我们的方法是可行的，以评估的过程中，干细胞衍生的神经元整合到一个主机神经元网络。
Stem cell transplantation is a promising therapy to treat neurodegenerative disorders, and a number ofin vitromodels have been developed for studying interactions between grafted neurons and the host neuronal network to promote drug discovery. However, methods capable of evaluating the process by which stem cells integrate into the host neuronal network are lacking. In this study, we applied an axonal conduction-based analysis to a co-culture study of primary and differentiated neurons. Mouse cortical neurons and neuronal cells differentiated from P19 embryonal carcinoma cells, a model for early neural differentiation of pluripotent stem cells, were co-cultured in a microfabricated device. The somata of these cells were separated by the co-culture device, but their axons were able to elongate through microtunnels and then form synaptic contacts. Propagating action potentials were recorded from these axons by microelectrodes embedded at the bottom of the microtunnels and sorted into clusters representing individual axons. While the number of axons of cortical neurons increased until 14 daysin vitroand then decreased, those of P19 neurons increased throughout the culture period. Network burst analysis showed that P19 neurons participated in approximately 80 % of the bursting activity after 14 daysin vitro. Interestingly, the axonal conduction delay of P19 neurons was significantly greater than that of cortical neurons, suggesting that there are some physiological differences in their axons. These results suggest that our method is feasible to evaluate the process by which stem cell-derived neurons integrate into a host neuronal network.
一种快速有效的 P19 胚胎癌细胞系神经元诱导方法
影响因子：
3
作者：
Y. Nakayama;A. Wada;Rei Inoue;Kazuya Terasawa;I. Kimura;Naosuke Nakamura;A. Kurosaka
通讯作者：
A. Kurosaka
兴奋性和抑制性层间新皮质轴突的比较特性
影响因子：
3.3
作者：
D. Shlosberg;Yasmin Abu;Y. Amitai
通讯作者：
Y. Amitai
神经元活动同步促进早期发育中大鼠新皮质神经元的存活
影响因子：
3.4
作者：
T. Voigt;H. Baier;A. D. Lima
通讯作者：
A. D. Lima