Isolation of inflammatory cells from rat brain tissue after stroke.

Karoline Möller, Tobias Stahl, Johannes Boltze, Daniel-Christoph Wagner
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引用次数: 14

Abstract

The pathophysiology of sterile inflammation following focal ischemic stroke is complex and not fully understood, but there is growing evidence that it offers several therapeutic options beyond the hitherto existing treatment strategies. The identification and quantification of infiltrating inflammatory cells in animal models of stroke is crucial both for understanding post-stroke inflammation and for drug target identification. Multicolor flow cytometry plays an important role in determining subtypes and quantity of leukocytes that infiltrate the brain tissue after stroke. Until now, most investigations have been performed in mice, most likely due to a significantly broader spectrum of disposable antibodies and available knockout models. Here, we introduce a specific and reproducible method to isolate leukocytes from rat brain specimen in the context of brain ischemia to ultimately allow multi-dimensional flow cytometric characterization and further downstream methods such as cell-subtype sorting and molecular biological approaches.

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脑卒中后大鼠脑组织炎症细胞的分离。
局灶性缺血性脑卒中后无菌性炎症的病理生理学是复杂的,尚未完全了解,但越来越多的证据表明,它提供了几种迄今为止现有的治疗策略之外的治疗选择。脑卒中动物模型中浸润炎性细胞的鉴定和定量对于理解脑卒中后炎症和药物靶点鉴定都至关重要。多色流式细胞术在确定脑卒中后脑组织浸润白细胞的亚型和数量方面起着重要作用。到目前为止,大多数研究都是在小鼠身上进行的,很可能是因为一次性抗体的范围更广,而且有可用的敲除模型。在此,我们介绍了一种在脑缺血情况下从大鼠脑标本中分离白细胞的特异性和可重复性方法,最终实现了多维流式细胞术表征和进一步的下游方法,如细胞亚型分类和分子生物学方法。
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