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Isolation, Purification, and Culture of Embryonic Melanoblasts from Green Fluorescent Protein–expressing Reporter Mice

In this article, we provide a method to isolate embryonic melanoblasts from reporter mouse strains. The mice from which these cells are isolated are bred into the ROSA26(mT/mG) reporter background, which results in green fluorescent protein (GFP) expression in the targeted melanoblast population. Th...

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Detalles Bibliográficos
Autores principales: Crawford, Melissa, Barr, Kevin, Dagnino, Lina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Bio-Protocol 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10501914/
https://www.ncbi.nlm.nih.gov/pubmed/37719067
http://dx.doi.org/10.21769/BioProtoc.4805
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author Crawford, Melissa
Barr, Kevin
Dagnino, Lina
author_facet Crawford, Melissa
Barr, Kevin
Dagnino, Lina
author_sort Crawford, Melissa
collection PubMed
description In this article, we provide a method to isolate embryonic melanoblasts from reporter mouse strains. The mice from which these cells are isolated are bred into the ROSA26(mT/mG) reporter background, which results in green fluorescent protein (GFP) expression in the targeted melanoblast population. These cells are isolated and purified by fluorescence-activated cell sorting using GFP fluorescence. We also provide a method to culture the purified melanoblasts for further analysis. This method yields > 99% purity melanoblasts specifically targeted, and can be used for a variety of studies, including gene expression, clonogenic experiments, and biological assays, such as viability, capacity for directional migration, or differentiation into melanin-producing melanocytic cells.
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spelling pubmed-105019142023-09-16 Isolation, Purification, and Culture of Embryonic Melanoblasts from Green Fluorescent Protein–expressing Reporter Mice Crawford, Melissa Barr, Kevin Dagnino, Lina Bio Protoc Methods Article In this article, we provide a method to isolate embryonic melanoblasts from reporter mouse strains. The mice from which these cells are isolated are bred into the ROSA26(mT/mG) reporter background, which results in green fluorescent protein (GFP) expression in the targeted melanoblast population. These cells are isolated and purified by fluorescence-activated cell sorting using GFP fluorescence. We also provide a method to culture the purified melanoblasts for further analysis. This method yields > 99% purity melanoblasts specifically targeted, and can be used for a variety of studies, including gene expression, clonogenic experiments, and biological assays, such as viability, capacity for directional migration, or differentiation into melanin-producing melanocytic cells. Bio-Protocol 2023-09-05 /pmc/articles/PMC10501914/ /pubmed/37719067 http://dx.doi.org/10.21769/BioProtoc.4805 Text en ©Copyright : © 2023 The Authors; This is an open access article under the CC BY-NC license https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the CC BY-NC license (https://creativecommons.org/licenses/by-nc/4.0/).
spellingShingle Methods Article
Crawford, Melissa
Barr, Kevin
Dagnino, Lina
Isolation, Purification, and Culture of Embryonic Melanoblasts from Green Fluorescent Protein–expressing Reporter Mice
title Isolation, Purification, and Culture of Embryonic Melanoblasts from Green Fluorescent Protein–expressing Reporter Mice
title_full Isolation, Purification, and Culture of Embryonic Melanoblasts from Green Fluorescent Protein–expressing Reporter Mice
title_fullStr Isolation, Purification, and Culture of Embryonic Melanoblasts from Green Fluorescent Protein–expressing Reporter Mice
title_full_unstemmed Isolation, Purification, and Culture of Embryonic Melanoblasts from Green Fluorescent Protein–expressing Reporter Mice
title_short Isolation, Purification, and Culture of Embryonic Melanoblasts from Green Fluorescent Protein–expressing Reporter Mice
title_sort isolation, purification, and culture of embryonic melanoblasts from green fluorescent protein–expressing reporter mice
topic Methods Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10501914/
https://www.ncbi.nlm.nih.gov/pubmed/37719067
http://dx.doi.org/10.21769/BioProtoc.4805
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