Sequential Isolation of Microglia and Astrocytes from Young and Aged Adult Mouse Brains for Downstream Transcriptomic Analysis.
Sequential Isolation of Microglia and Astrocytes from Young and Aged Adult Mouse Brains for Downstream Transcriptomic Analysis.
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DOI：
10.3390/mps5050077
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发表时间：
2022-09-27
影响因子：
2.4
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中科院分区：
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随着年龄的增长，大脑更容易受到损伤和神经退行性疾病的影响，但其机制在很大程度上是未知的。目前有证据表明，由常驻脑星形胶质细胞和小胶质细胞群介导的神经炎症是炎症反应产生的关键参与者，可能影响与年龄相关的过程和神经退行性变的开始/进展。因此，单独和集体靶向这些细胞类型可能有助于开发新的疾病修饰疗法。我们已经优化和表征了一种方案，用于有效地从年轻和老年小鼠的成年小鼠大脑中分离小胶质细胞和星形胶质细胞。我们展示了一种利用磁珠技术对这些免疫细胞进行顺序分离的技术，该技术经过优化，可以提高产量，并限制下游转录组学应用（包括rna测序管道）中潜在的伪影。该技术用途广泛，成本效益高，在相同的生物学背景下研究反应可靠，同时有利于减少评估正常和年龄相关病理条件下细胞反应所需的小鼠数量。
In aging, the brain is more vulnerable to injury and neurodegenerative disease, but the mechanisms responsible are largely unknown. Evidence now suggests that neuroinflammation, mediated by resident brain astrocyte and microglia populations, are key players in the generation of inflammatory responses and may influence both age related processes and the initiation/progression of neurodegeneration. Consequently, targeting these cell types individually and collectively may aid in the development of novel disease-modifying therapies. We have optimized and characterized a protocol for the effective sequential isolation of both microglia and astrocytes from the adult mouse brain in young and aged mice. We demonstrate a technique for the sequential isolation of these immune cells by using magnetic beads technology, optimized to increase yield and limit potential artifacts in downstream transcriptomic applications, including RNA-sequencing pipelines. This technique is versatile, cost-effective, and reliable for the study of responses within the same biological context, simultaneously being advantageous in reducing mice numbers required to assess cellular responses in normal and age-related pathological conditions.