Differentiation and characterization of neurons derived from rat iPSCs.
Differentiation and characterization of neurons derived from rat iPSCs.
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DOI：
10.1016/j.jneumeth.2020.108693
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发表时间：
2020-05-15
影响因子：
3
通讯作者：
Purcell EK
中科院分区：
文献类型：
--
作者：
Setien MB;Smith KR;Howard K;Williams K;Suhr ST;Purcell EK
诱导多能干细胞(IPSCs)可能是修复神经系统疾病或创伤所致损伤的神经细胞的有利来源。此外，他们很有希望开发模型来研究神经退行性疾病的潜在机制。虽然在小鼠中已有成功的IPSCs神经分化的报道，但详细描述大鼠IPSCs神经细胞生成的方案相对有限，通过操纵细胞培养方法来优化它们仍未被探索。在这里，我们描述和比较了四种不同的、常用的底物对大鼠ipSC(RiPSC)来源的神经前体细胞分化的影响。我们的方法是使用底物包被作为一种方法来丰富分化的rPSCs，以获得所需的形态和成熟度的神经元亚型。我们使用电生理学、免疫荧光染色和Sholl分析相结合的方法来表征在每种底物上产生的细胞在9天的时间过程中的特征。细胞培养底物的表面涂层影响分化神经元的极性和树枝。聚鸟氨酸-层粘连蛋白涂层促进神经元的树枝和成熟，而Geltrex有利于表现出功能不成熟迹象的双极细胞。多聚-D-赖氨酸底物与有限的轴突生长和树枝形成有关。明胶是最不利于细胞生长和分化的底物。以前已经有大鼠神经前体细胞的来源；然而，我们使用底物涂层来影响形态和电成熟的方法还没有被探索过。可以选择底物涂层来丰富分化的rPSCs，以获得不同的神经元形态。
Induced pluripotent stem cells (iPSCs) may be an advantageous source of neuronal cells to repair damage due to neurological disorders or trauma. Additionally, they are promising candidates to develop models to study underlying mechanisms of neurodegenerative diseases. While successful neural differentiation of iPSCs has been reported in mice, protocols detailing the generation of neural cells from rat iPSCs are relatively limited, and their optimization by manipulating cell culture methods has remained unexplored. Here, we describe and compare the effects of four distinct, commonly used substrates on the neuronal differentiation of rat iPSC (riPSC) derived-neural progenitor cells. Our approach is to use substrate coating as a method to enrich differentiated riPSCs for neuronal subtypes with the desired morphology and maturity. We use a combination of electrophysiology, immunofluorescence staining, and Sholl analysis to characterize the cells generated on each substrate over a nine-day time course. The surface coating presented by the cell culture substrate influences the polarity and arborization of differentiating neurons. Polyornithine-laminin coating promoted neuronal arborization and maturation, while Geltrex favored bipolar cells which displayed indicators of functional immaturity. Poly-D-lysine substrate was associated with limited neurite outgrowth and arborization. Gelatin was the least favorable substrate for the growth and differentiation of our cells. Rat-derived neural progenitor cells have been previously derived; however, our methods to use substrate coatings to influence morphological and electrical maturity have not been explored previously. Substrate coatings can be selected to enrich differentiated riPSCs for distinctive neuronal morphologies.
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影响因子：
25
作者：
Johnson, Stuart L.;Eckrich, Tobias;Kuhn, Stephanie;Zampini, Valeria;Franz, Christoph;Ranatunga, Kishani M.;Roberts, Terri P.;Masetto, Sergio;Knipper, Marlies;Kros, Corne J.;Marcotti, Walter
通讯作者：
Marcotti, Walter
DOI：
10.1073/pnas.0910012107
发表时间：
2010-03-02
影响因子：
11.1
作者：
Hu, Bao-Yang;Weick, Jason P.;Zhang, Su-Chun
通讯作者：
Zhang, Su-Chun
聚-L-鸟氨酸通过 ERK 信号通路促进神经干/祖细胞的优先分化
影响因子：
4.6
作者：
Ge H;Tan L;Wu P;Yin Y;Liu X;Meng H;Cui G;Wu N;Lin J;Hu R;Feng H
通讯作者：
Feng H
影响因子：
3.7
作者：
Hamanaka S;Yamaguchi T;Kobayashi T;Kato-Itoh M;Yamazaki S;Sato H;Umino A;Wakiyama Y;Arai M;Sanbo M;Hirabayashi M;Nakauchi H
通讯作者：
Nakauchi H
影响因子：
2.4
作者：
Illes S;Theiss S;Hartung HP;Siebler M;Dihné M
通讯作者：
Dihné M