通过分段传代纯化原始神经祖细胞以产生富含星形胶质细胞的脑类器官的方法。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2025-10-13 DOI:10.1016/j.xpro.2025.104142

Kushal Aluru, Rui Dang, Mengmeng Jin, Peng Jiang

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引用次数: 0

摘要

人类多能干细胞（hPSC）衍生的原始神经祖细胞（pnpc）和脑类器官为研究人类大脑发育提供了一个强大的平台。在这里，我们提出了一个分数传递协议，产生几乎纯PAX6+/SOX2+ pnpc。我们描述了神经上皮分化、纯化和生产富含星形胶质细胞的类器官的步骤。然后详细介绍下游分析的程序。类器官含有超过15%的星形胶质细胞和成熟的神经元，提供了一个可扩展和可复制的系统来模拟人类胶质-神经元在发育和疾病中的相互作用。有关本协议使用和执行的完整细节，请参阅Dang等人1。

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Protocol for purifying primitive neural progenitors by fractional passaging to generate astroglia-enriched cerebral organoid.

Human pluripotent stem cell (hPSC)-derived primitive neural progenitor cells (pNPCs) and cerebral organoids provide a powerful platform for studying human brain development. Here, we present a fractional passaging protocol that yields nearly pure PAX6⁺/SOX2⁺ pNPCs. We describe steps for neuroepithelial differentiation, purification, and production of astroglia-enriched organoids. We then detail procedures for downstream analysis. The organoids contain over 15% astrocytes alongside maturing neurons, providing a scalable and reproducible system to model human glial-neuronal interactions in development and disease. For complete details on the use and execution of this protocol, please refer to Dang et al.¹.

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