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Normal and Malignant Muscle Cell Transplantation into Immune Compromised Adult Zebrafish

Zebrafish have become a powerful tool for assessing development, regeneration, and cancer. More recently, allograft cell transplantation protocols have been developed that permit engraftment of normal and malignant cells into irradiated, syngeneic, and immune compromised adult zebrafish. These model...

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Autores principales: Tenente, Inês M., Tang, Qin, Moore, John C., Langenau, David M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4354485/
https://www.ncbi.nlm.nih.gov/pubmed/25591079
http://dx.doi.org/10.3791/52597
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author Tenente, Inês M.
Tang, Qin
Moore, John C.
Langenau, David M.
author_facet Tenente, Inês M.
Tang, Qin
Moore, John C.
Langenau, David M.
author_sort Tenente, Inês M.
collection PubMed
description Zebrafish have become a powerful tool for assessing development, regeneration, and cancer. More recently, allograft cell transplantation protocols have been developed that permit engraftment of normal and malignant cells into irradiated, syngeneic, and immune compromised adult zebrafish. These models when coupled with optimized cell transplantation protocols allow for the rapid assessment of stem cell function, regeneration following injury, and cancer. Here, we present a method for cell transplantation of zebrafish adult skeletal muscle and embryonal rhabdomyosarcoma (ERMS), a pediatric sarcoma that shares features with embryonic muscle, into immune compromised adult rag2(E450fs )homozygous mutant zebrafish. Importantly, these animals lack T cells and have reduced B cell function, facilitating engraftment of a wide range of tissues from unrelated donor animals. Our optimized protocols show that fluorescently labeled muscle cell preparations from α-actin-RFP transgenic zebrafish engraft robustly when implanted into the dorsal musculature of rag2 homozygous mutant fish. We also demonstrate engraftment of fluorescent-transgenic ERMS where fluorescence is confined to cells based on differentiation status. Specifically, ERMS were created in AB-strain myf5-GFP; mylpfa-mCherry double transgenic animals and tumors injected into the peritoneum of adult immune compromised fish. The utility of these protocols extends to engraftment of a wide range of normal and malignant donor cells that can be implanted into dorsal musculature or peritoneum of adult zebrafish.
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spelling pubmed-43544852015-03-12 Normal and Malignant Muscle Cell Transplantation into Immune Compromised Adult Zebrafish Tenente, Inês M. Tang, Qin Moore, John C. Langenau, David M. J Vis Exp Immunology Zebrafish have become a powerful tool for assessing development, regeneration, and cancer. More recently, allograft cell transplantation protocols have been developed that permit engraftment of normal and malignant cells into irradiated, syngeneic, and immune compromised adult zebrafish. These models when coupled with optimized cell transplantation protocols allow for the rapid assessment of stem cell function, regeneration following injury, and cancer. Here, we present a method for cell transplantation of zebrafish adult skeletal muscle and embryonal rhabdomyosarcoma (ERMS), a pediatric sarcoma that shares features with embryonic muscle, into immune compromised adult rag2(E450fs )homozygous mutant zebrafish. Importantly, these animals lack T cells and have reduced B cell function, facilitating engraftment of a wide range of tissues from unrelated donor animals. Our optimized protocols show that fluorescently labeled muscle cell preparations from α-actin-RFP transgenic zebrafish engraft robustly when implanted into the dorsal musculature of rag2 homozygous mutant fish. We also demonstrate engraftment of fluorescent-transgenic ERMS where fluorescence is confined to cells based on differentiation status. Specifically, ERMS were created in AB-strain myf5-GFP; mylpfa-mCherry double transgenic animals and tumors injected into the peritoneum of adult immune compromised fish. The utility of these protocols extends to engraftment of a wide range of normal and malignant donor cells that can be implanted into dorsal musculature or peritoneum of adult zebrafish. MyJove Corporation 2014-12-26 /pmc/articles/PMC4354485/ /pubmed/25591079 http://dx.doi.org/10.3791/52597 Text en Copyright © 2014, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Immunology
Tenente, Inês M.
Tang, Qin
Moore, John C.
Langenau, David M.
Normal and Malignant Muscle Cell Transplantation into Immune Compromised Adult Zebrafish
title Normal and Malignant Muscle Cell Transplantation into Immune Compromised Adult Zebrafish
title_full Normal and Malignant Muscle Cell Transplantation into Immune Compromised Adult Zebrafish
title_fullStr Normal and Malignant Muscle Cell Transplantation into Immune Compromised Adult Zebrafish
title_full_unstemmed Normal and Malignant Muscle Cell Transplantation into Immune Compromised Adult Zebrafish
title_short Normal and Malignant Muscle Cell Transplantation into Immune Compromised Adult Zebrafish
title_sort normal and malignant muscle cell transplantation into immune compromised adult zebrafish
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4354485/
https://www.ncbi.nlm.nih.gov/pubmed/25591079
http://dx.doi.org/10.3791/52597
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