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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2023
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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 |
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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 |
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