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Isolation of single cells from individual mouse ovaries for flow cytometry and functional analysis

Here, we present a validated workflow to isolate sufficient viable single ovary cells from a single mouse without the need to pool from several mice. We provide steps essential for estrous staging, ovary harvesting and dissociation, ovary cell staining, data collection, and analysis. Our approach al...

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Detalles Bibliográficos
Autores principales: Bafor, Enitome E., Martin, Toni, Carrell, Jeffrey, Karwan, Megan, Kimmel, Adrienne E., Omogiade, Uyi.G., Sanford, Michael, Young, Howard A., Valencia, Julio C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10679862/
https://www.ncbi.nlm.nih.gov/pubmed/37963022
http://dx.doi.org/10.1016/j.xpro.2023.102710
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author Bafor, Enitome E.
Martin, Toni
Carrell, Jeffrey
Karwan, Megan
Kimmel, Adrienne E.
Omogiade, Uyi.G.
Sanford, Michael
Young, Howard A.
Valencia, Julio C.
author_facet Bafor, Enitome E.
Martin, Toni
Carrell, Jeffrey
Karwan, Megan
Kimmel, Adrienne E.
Omogiade, Uyi.G.
Sanford, Michael
Young, Howard A.
Valencia, Julio C.
author_sort Bafor, Enitome E.
collection PubMed
description Here, we present a validated workflow to isolate sufficient viable single ovary cells from a single mouse without the need to pool from several mice. We provide steps essential for estrous staging, ovary harvesting and dissociation, ovary cell staining, data collection, and analysis. Our approach allows the use of these single-cell suspensions for flow sorting, flow cytometry analysis, or functional in vitro assays. Importantly, our protocol is designed to maximize the isolation of immune cells, including T cell subsets.
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spelling pubmed-106798622023-11-14 Isolation of single cells from individual mouse ovaries for flow cytometry and functional analysis Bafor, Enitome E. Martin, Toni Carrell, Jeffrey Karwan, Megan Kimmel, Adrienne E. Omogiade, Uyi.G. Sanford, Michael Young, Howard A. Valencia, Julio C. STAR Protoc Protocol Here, we present a validated workflow to isolate sufficient viable single ovary cells from a single mouse without the need to pool from several mice. We provide steps essential for estrous staging, ovary harvesting and dissociation, ovary cell staining, data collection, and analysis. Our approach allows the use of these single-cell suspensions for flow sorting, flow cytometry analysis, or functional in vitro assays. Importantly, our protocol is designed to maximize the isolation of immune cells, including T cell subsets. Elsevier 2023-11-14 /pmc/articles/PMC10679862/ /pubmed/37963022 http://dx.doi.org/10.1016/j.xpro.2023.102710 Text en © 2023. 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
Bafor, Enitome E.
Martin, Toni
Carrell, Jeffrey
Karwan, Megan
Kimmel, Adrienne E.
Omogiade, Uyi.G.
Sanford, Michael
Young, Howard A.
Valencia, Julio C.
Isolation of single cells from individual mouse ovaries for flow cytometry and functional analysis
title Isolation of single cells from individual mouse ovaries for flow cytometry and functional analysis
title_full Isolation of single cells from individual mouse ovaries for flow cytometry and functional analysis
title_fullStr Isolation of single cells from individual mouse ovaries for flow cytometry and functional analysis
title_full_unstemmed Isolation of single cells from individual mouse ovaries for flow cytometry and functional analysis
title_short Isolation of single cells from individual mouse ovaries for flow cytometry and functional analysis
title_sort isolation of single cells from individual mouse ovaries for flow cytometry and functional analysis
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10679862/
https://www.ncbi.nlm.nih.gov/pubmed/37963022
http://dx.doi.org/10.1016/j.xpro.2023.102710
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