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

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 c...

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Detalles Bibliográficos
Autores principales: Chowdhury, Shanin, Fried, Kaela Drzewiecki, Iwakiri, Yasuko, Brancale, Joseph, Vilarinho, Sílvia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10400957/
https://www.ncbi.nlm.nih.gov/pubmed/37515764
http://dx.doi.org/10.1016/j.xpro.2023.102480
Descripción
Sumario: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).(1)