Cargando…
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...
Autores principales: | , , |
---|---|
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 |
_version_ | 1785106209202765824 |
---|---|
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. |
format | Online Article Text |
id | pubmed-10501914 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Bio-Protocol |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT crawfordmelissa isolationpurificationandcultureofembryonicmelanoblastsfromgreenfluorescentproteinexpressingreportermice AT barrkevin isolationpurificationandcultureofembryonicmelanoblastsfromgreenfluorescentproteinexpressingreportermice AT dagninolina isolationpurificationandcultureofembryonicmelanoblastsfromgreenfluorescentproteinexpressingreportermice |