Generation and assembly of human brain region-specific three-dimensional cultures.
Generation and assembly of human brain region-specific three-dimensional cultures.
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DOI：
10.1038/s41596-018-0032-7
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发表时间：
2018-09
期刊：
影响因子：
14.8
通讯作者：
Pașca SP
中科院分区：
文献类型：
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作者：
Sloan SA;Andersen J;Pașca AM;Birey F;Pașca SP
生成特定区域的3D模型来研究人类大脑发育的能力为理解健康个体和患者的神经系统提供了巨大的希望。在该方案中，我们描述了如何从人多能干细胞（hPSC）生成和组装亚结构域特异性前脑球状体，也称为脑区域特异性类器官。我们描述了如何图案的神经球体对背侧前脑或腹侧前脑的命运，建立人类皮质球体（hCS）或，人类的subpallial球体（hSS），分别。我们还描述了如何在体外联合收割机的神经球体组装前脑胶质细胞，重演的相互作用的多巴胺能和GABA能神经元在体内看到。星形胶质细胞也存在于人前脑特异性球体中，并且当前脑球体长期培养时，这些星形胶质细胞经历成熟。从hPSC的神经球体的初始生成发生在不到一周内，在随后的5周内发生区域图案化。在初始成熟阶段之后，脑区域特异性球状体适合于各种测定，包括活细胞成像、钙信号传导、电生理学、细胞纯化、单细胞转录组学和免疫组织化学。一旦产生，前脑球状体也可以在培养中成熟超过24个月。
The ability to generate region-specific 3D models to study human brain development offers great promise for understanding the nervous system in both healthy individuals and patients. In this protocol we describe how to generate and assemble subdomain-specific forebrain spheroids, also known as brain-region specific organoids, from human pluripotent stem cells (hPSCs). We describe how to pattern the neural spheroids towards either a dorsal forebrain or ventral forebrain fate, establishing human cortical spheroids (hCS) or, human subpallial spheroids (hSS), respectively. We also describe how to combine the neural spheroids in vitro to assemble forebrain assembloids that recapitulate the interactions of glutamatergic and GABAergic neurons seen in vivo. Astrocytes are also present in the human forebrain-specific spheroids and these undergo maturation when the forebrain spheroids are cultured long-term. The initial generation of neural spheroids from hPSCs occurs in less than one week, with regional patterning occurring over the subsequent 5 weeks. After the initial maturation stage, brain region-specific spheroids are amenable to a variety of assays, including live cell imaging, calcium signaling, electrophysiology, cell purification, single cell transcriptomics, and immunohistochemistry. Once generated, forebrain spheroids can also be matured for over 24 months in culture.
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DOI：
10.1126/science.aaf7073
发表时间：
2016-10-07
期刊：
Science (New York, N.Y.)
影响因子：
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作者：
Paredes MF;James D;Gil-Perotin S;Kim H;Cotter JA;Ng C;Sandoval K;Rowitch DH;Xu D;McQuillen PS;Garcia-Verdugo JM;Huang EJ;Alvarez-Buylla A
通讯作者：
Alvarez-Buylla A
影响因子：
64.5
作者：
Mariani J;Coppola G;Zhang P;Abyzov A;Provini L;Tomasini L;Amenduni M;Szekely A;Palejev D;Wilson M;Gerstein M;Grigorenko EL;Chawarska K;Pelphrey KA;Howe JR;Vaccarino FM
通讯作者：
Vaccarino FM
影响因子：
11
作者：
Brennand, K. J.;Simone, A.;Tran, N.;Gage, F. H.
通讯作者：
Gage, F. H.
影响因子：
64.5
作者：
Dolmetsch R;Geschwind DH
通讯作者：
Geschwind DH
影响因子：
64.8
作者：
通讯作者：
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