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富集、纯化和细胞离心法分离小鼠肝内皮细胞的方案。

Protocol for enrichment, purification, and cytocentrifugation of mouse liver endothelial cells.

机构信息

Department of Internal Medicine, Section of Digestive Diseases, Yale School of Medicine, New Haven, CT 06519, USA; Department of Pathology, Yale School of Medicine, New Haven, CT 06519, USA.

Department of Internal Medicine, Section of Digestive Diseases, Yale School of Medicine, New Haven, CT 06519, USA.

出版信息

STAR Protoc. 2023 Sep 15;4(3):102480. doi: 10.1016/j.xpro.2023.102480. Epub 2023 Jul 28.

Abstract

Liver endothelial cells (LECs) are critical in maintaining liver homeostasis. To understand the mechanistic processes occurring in these cells, high-quality isolation protocols must be in place. Here, we present a protocol for LEC enrichment, subsequent LEC purification using fluorescence-assisted cell sorting, and cytocentrifugation of sorted LECs for imaging. We describe steps for isolation of LEC-enriched population from mouse livers, immunolabeling and sorting, and cytospin and immunostaining. We then mention procedures for downstream analysis. For complete details on the use and execution of this protocol, please refer to Drzewiecki et al. (2021)..

摘要

肝脏内皮细胞(LECs)对于维持肝脏内环境稳定至关重要。为了了解这些细胞中发生的机制过程,必须要有高质量的分离方案。在这里,我们提出了一种从小鼠肝脏中富集 LEC 的方案,然后使用荧光激活细胞分选对 LEC 进行纯化,并对分选的 LEC 进行细胞离心以进行成像。我们描述了从小鼠肝脏中分离富含 LEC 群体、免疫标记和分选以及细胞离心和免疫染色的步骤。然后,我们提到了下游分析的程序。有关此方案使用和执行的完整详细信息,请参考 Drzewiecki 等人。(2021 年)。

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