Cargando…

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...

Descripción completa

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
_version_ 1785084554068885504
author Chowdhury, Shanin
Fried, Kaela Drzewiecki
Iwakiri, Yasuko
Brancale, Joseph
Vilarinho, Sílvia
author_facet Chowdhury, Shanin
Fried, Kaela Drzewiecki
Iwakiri, Yasuko
Brancale, Joseph
Vilarinho, Sílvia
author_sort Chowdhury, Shanin
collection PubMed
description 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)
format Online
Article
Text
id pubmed-10400957
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-104009572023-08-05 Protocol for enrichment, purification, and cytocentrifugation of mouse liver endothelial cells Chowdhury, Shanin Fried, Kaela Drzewiecki Iwakiri, Yasuko Brancale, Joseph Vilarinho, Sílvia STAR Protoc Protocol 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) Elsevier 2023-07-28 /pmc/articles/PMC10400957/ /pubmed/37515764 http://dx.doi.org/10.1016/j.xpro.2023.102480 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Chowdhury, Shanin
Fried, Kaela Drzewiecki
Iwakiri, Yasuko
Brancale, Joseph
Vilarinho, Sílvia
Protocol for enrichment, purification, and cytocentrifugation of mouse liver endothelial cells
title Protocol for enrichment, purification, and cytocentrifugation of mouse liver endothelial cells
title_full Protocol for enrichment, purification, and cytocentrifugation of mouse liver endothelial cells
title_fullStr Protocol for enrichment, purification, and cytocentrifugation of mouse liver endothelial cells
title_full_unstemmed Protocol for enrichment, purification, and cytocentrifugation of mouse liver endothelial cells
title_short Protocol for enrichment, purification, and cytocentrifugation of mouse liver endothelial cells
title_sort protocol for enrichment, purification, and cytocentrifugation of mouse liver endothelial cells
topic Protocol
url 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
work_keys_str_mv AT chowdhuryshanin protocolforenrichmentpurificationandcytocentrifugationofmouseliverendothelialcells
AT friedkaeladrzewiecki protocolforenrichmentpurificationandcytocentrifugationofmouseliverendothelialcells
AT iwakiriyasuko protocolforenrichmentpurificationandcytocentrifugationofmouseliverendothelialcells
AT brancalejoseph protocolforenrichmentpurificationandcytocentrifugationofmouseliverendothelialcells
AT vilarinhosilvia protocolforenrichmentpurificationandcytocentrifugationofmouseliverendothelialcells