Establishment of an Electrophysiological Platform for Modeling ALS with Regionally-Specific Human Pluripotent Stem Cell-Derived Astrocytes and Neurons.
Establishment of an Electrophysiological Platform for Modeling ALS with Regionally-Specific Human Pluripotent Stem Cell-Derived Astrocytes and Neurons.
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DOI：
10.3791/62726
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发表时间：
2021-08-26
影响因子：
1.2
通讯作者：
Maragakis, Nicholas J.
中科院分区：
文献类型：
--
作者：
Taga, Arens;Habela, Christa W.;Johns, Alexandra;Liu, Shiyu;O'Brien, Mollie;Maragakis, Nicholas J.
人多能干细胞来源的星形胶质细胞(hiPSC-A)和神经元(hiPSC-N)为体外模拟肌萎缩侧索硬化症(ALS)的病理生理机制提供了有力的工具。多电极阵列(MEA)记录是一种记录大量神经元的电场电位并分析随时间变化的网络活动的方法。先前的研究表明，与没有hIPSC-A的培养或在啮齿动物星形胶质细胞存在的情况下相比，使用促进脊髓星形胶质细胞表型的技术分化的hiPSC-A的存在可以改善局部特定的脊髓hiPSC运动神经元(MN)的成熟和电生理活性。本文描述了一种将脊髓HIPSC-A与HIPSC-MN共培养并使用MEA记录记录电生理活动的方法。虽然这里描述的分化方案是特定于脊髓区域特有的星形胶质细胞和神经元，但共培养平台可以应用于通过特定于其他命运的技术分化的星形胶质细胞和神经元，包括皮质HiPSC-A和HiPSC-N。这些方案旨在提供一种电生理测试，以了解神经胶质细胞与神经元的相互作用，并为测试具有治疗潜力的药物提供一个平台。
Human pluripotent stem cell-derived astrocytes (hiPSC-A) and neurons (hiPSC-N) provide a powerful tool for modeling Amyotrophic Lateral Sclerosis (ALS) pathophysiology in vitro. Multi-electrode array (MEA) recordings are a means to record electrical field potentials from large populations of neurons and analyze network activity over time. It was previously demonstrated that the presence of hiPSC-A that are differentiated using techniques to promote a spinal cord astrocyte phenotype improved maturation and electrophysiological activity of regionally specific spinal cord hiPSC-motor neurons (MN) when compared to those cultured without hiPSC-A or in the presence of rodent astrocytes. Described here is a method to co-culture spinal cord hiPSC-A with hiPSC-MN and record electrophysiological activity using MEA recordings. While the differentiation protocols described here are particular to astrocytes and neurons that are regionally specific to the spinal cord, the co-culturing platform can be applied to astrocytes and neurons differentiated with techniques specific to other fates, including cortical hiPSC-A and hiPSC-N. These protocols aim to provide an electrophysiological assay to inform about glia-neuron interactions and provide a platform for testing drugs with therapeutic potential in ALS.
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影响因子：
8.8
作者：
Wainger BJ;Kiskinis E;Mellin C;Wiskow O;Han SS;Sandoe J;Perez NP;Williams LA;Lee S;Boulting G;Berry JD;Brown RH Jr;Cudkowicz ME;Bean BP;Eggan K;Woolf CJ
通讯作者：
Woolf CJ
影响因子：
8.8
作者：
Roybon L;Lamas NJ;Garcia AD;Yang EJ;Sattler R;Lewis VJ;Kim YA;Kachel CA;Rothstein JD;Przedborski S;Wichterle H;Henderson CE
通讯作者：
Henderson CE
影响因子：
4.6
作者：
Odawara A;Matsuda N;Ishibashi Y;Yokoi R;Suzuki I
通讯作者：
Suzuki I
影响因子：
3.6
作者：
Shimojo D;Onodera K;Doi-Torii Y;Ishihara Y;Hattori C;Miwa Y;Tanaka S;Okada R;Ohyama M;Shoji M;Nakanishi A;Doyu M;Okano H;Okada Y
通讯作者：
Okada Y
影响因子：
6.2
作者：
Almad AA;Doreswamy A;Gross SK;Richard JP;Huo Y;Haughey N;Maragakis NJ
通讯作者：
Maragakis NJ