Laminin α5 substrates promote survival, network formation and functional development of human pluripotent stem cell-derived neurons in vitro
Laminin α5 substrates promote survival, network formation and functional development of human pluripotent stem cell-derived neurons in vitro
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DOI：
10.1016/j.scr.2017.09.002
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发表时间：
2017-10-01
影响因子：
1.2
通讯作者：
Narkilahti, Susanna
中科院分区：
文献类型：
--
作者：
Hyysalo, Anu;Ristola, Mervi;Narkilahti, Susanna
层粘连蛋白是细胞外基质(ECM)中的主要蛋白质组之一，体内特定的层粘连蛋白亚型对中枢神经系统的神经功能至关重要。在本研究中，我们比较了重组人层粘连蛋白亚型(LN211、LN332、LN411、LN511和LN521)和层粘连蛋白亚型片段(LN511E8)在体外培养的人多能干细胞(HPSC)来源的神经元。通过相位对比成像、活性染色和免疫细胞化学检测，我们发现含有α5-链的层粘连蛋白底物对于神经元附着、活性和网络形成是重要的。基因表达分析表明，hPSC来源的神经元偏爱特定的层粘连蛋白亚型的分子机制可能与细胞外基质重塑和细胞黏附有关。重要的是，微电极阵列分析揭示了层粘连蛋白α5底物上电生理活性神经元的最广泛分布，表明神经元网络功能的最有效发展。这项研究表明，特定的层粘连蛋白α5底物为hPSC来源的神经元提供了一个可控的体外培养环境。这些底物不仅可以用于提高hPSC来源的功能性神经元的产量，用于疾病建模、毒理学研究和药物开发等体外应用，还可以用于生产临床级别的hPSC来源的细胞，用于再生医学应用。(C)2017年作者。由Elsevier B.V.出版。这是一篇CC BY-NC-ND许可证下的开放访问文章
Laminins are one of the major protein groups in the extracellular matrix (ECM) and specific laminin isoforms are crucial for neuronal functions in the central nervous system in vivo. In the present study, we compared recombinant human laminin isoforms (LN211, LN332, LN411, LN511, and LN521) and laminin isoform fragment (LN511E8) in in vitro cultures of human pluripotent stem cell (hPSC)-derived neurons. We showed that laminin substrates containing the alpha 5-chain are important for neuronal attachment, viability and network formation, as detected by phase contrast imaging, viability staining, and immunocytochemistry. Gene expression analysis showed that the molecular mechanisms involved in the preference of hPSC-derived neurons for specific laminin isoforms could be related to ECM remodeling and cell adhesion. Importantly, the microelectrodearray analysis revealed the widest distribution of electrophysiologically active neurons on laminin alpha 5 substrates, indicating most efficient development of neuronal network functionality. This study shows that specific laminin alpha 5 substrates provide a controlled in vitro culture environment for hPSC-derived neurons. These substrates can be utilized not only to enhance the production of functional hPSC-derived neurons for in vitro applications like disease modeling, toxicological studies, and drug discovery, but also for the production of clinical grade hPSC-derived cells for regenerative medicine applications. (C) 2017 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license