从单个小鼠幼崽中产生高度富集的性别特异性初级皮质星形胶质细胞的方案。

IF 1.4 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2026-10-03 DOI:10.1016/j.xpro.2026.104873

Sanarya Al-Jaf, En-Hung Ai, Julia A Wilson, Khaled S Abd-Elrahman

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引用次数: 0

摘要

在这里，我们提出了一种从单个新生小鼠幼崽中产生高度富集的初级皮层星形胶质细胞的方案。我们描述了机械分离皮质组织的步骤，在包被烧瓶中培养，并通过机械震动和选择性药理消耗小胶质细胞来去除少突胶质细胞祖细胞。然后，我们详细介绍了基于pcr的尾部活检DNA分析程序，以确定性别和基因型，从而从基因上定义星形细胞培养。该方案能够调查性别和基因型特异性机制，同时避免混合组织的混淆效应。

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Protocol for generating highly enriched sex-specific primary cortical astrocytes from individual mouse pups.

Here, we present a protocol for generating highly enriched primary cortical astrocytes from individual neonatal mouse pups. We describe steps for mechanically dissociating cortical tissue, culturing in coated flasks, and removing oligodendrocyte progenitor cells through mechanical shaking and selective pharmacological depletion of microglia. We then detail procedures for PCR-based analysis of tail-biopsy DNA to determine sex and genotype to genetically define astrocyte cultures. This protocol enables the investigation of sex- and genotype-specific mechanisms while avoiding the confounding effects of pooled tissue.

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