Flow Cytometric Characterization of Pluripotent Cell Protein Markers in Naïve, Formative, and Primed Pluripotent Stem Cells.

Animals Cell Differentiation Germ Layers Mice Mouse Embryonic Stem Cells Pluripotent Stem Cells
Embryonic stem cells Flow cytometry Formative pluripotency Naïve pluripotency Pluripotent stem cells Primed pluripotency mEpiSC

Journal

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969

Informations de publication

Date de publication:
2022
Historique:
entrez: 29 4 2022
pubmed: 30 4 2022
medline: 4 5 2022
Statut: ppublish

Résumé

Here we describe methodologies to characterize, delineate, and quantify pluripotent cells between naïve, formative, and primed pluripotent state mouse embryonic stem cell (mESCs) populations using flow cytometric analysis. This methodology can validate pluripotent states, sort individual cells of interest, and determine the efficiency of transitioning naïve mESCs to a primed-like state as mouse epiblast-like cells (mEpiLCs) and onto fully primed mouse epiblast stem cells (mEpiSCs). Quantification of the cell surface markers; SSEA1(CD15) and CD24 introduces an effective method of distinguishing individual cells from a population by their respective positioning in the pluripotent spectrum. Additionally, this protocol can be used to demarcate and sort cells via fluorescently activated cell sorting for downstream applications. Flow cytometric analysis within mESCs, mEpiLCs, and mEpiSCs can be efficiently completed using these optimized protocols.

Identifiants

DOI: 10.1007/978-1-0716-2281-0_8 PMID: 35486241
pubmed: 35486241
doi: 10.1007/978-1-0716-2281-0_8
doi:

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

81-92

Subventions

Organisme : CIHR
Pays : Canada

Informations de copyright

© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Références

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Auteurs

Joshua G Dierolf (JG)

Department of Physiology and Pharmacology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, Canada.

Kristin Chadwick (K)

Robarts Research Institute, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, Canada.

Courtney R Brooks (CR)

Department of Physiology and Pharmacology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, Canada.

Andrew J Watson (AJ)

Department of Physiology and Pharmacology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, Canada.
Department of Obstetrics and Gynecology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, Canada.
The Children's Health Research Institute (CHRI), Lawson Health Research Institute, London, ON, Canada.

Dean H Betts (DH)

Department of Physiology and Pharmacology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, Canada. dean.betts@schulich.uwo.ca.
Department of Obstetrics and Gynecology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, Canada. dean.betts@schulich.uwo.ca.
The Children's Health Research Institute (CHRI), Lawson Health Research Institute, London, ON, Canada. dean.betts@schulich.uwo.ca.

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Classifications MeSH

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questionsmedicales.fr › Phénomènes physiologiques cellulaires › Différenciation cellulaire › Flow Cytometric Characterization of...
questionsmedicales.fr › Structures de l'embryon › Feuillets embryonnaires › Flow Cytometric Characterization of...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Rodentia › Muridae › Murinae › Souris › Flow Cytometric Characterization of...
questionsmedicales.fr › Cellules › Cellules souches › Cellules souches pluripotentes › Cellules souches embryonnaires › Cellules souches embryonnaires de souris › Flow Cytometric Characterization of...
questionsmedicales.fr › Cellules › Cellules souches › Cellules souches pluripotentes › Flow Cytometric Characterization of...