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Retinoblastoma Cell Growth In Vitro and Tumor Formation In Ovo—Influence of Different Culture Conditions

Retinoblastoma (RB) is a primary intraocular malignancy in childhood. Relapses may develop and cause secondary cancers during later development. This study was set up to identify optimal cell culture conditions for RB cell growth in vitro and to optimize tumor growth in an in vivo model. RB cell lin...

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Autores principales: Doege, Annika, Steens, Rebecca, Dünker, Nicole, Busch, Maike Anna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8938814/
https://www.ncbi.nlm.nih.gov/pubmed/35314658
http://dx.doi.org/10.3390/mps5020021
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author Doege, Annika
Steens, Rebecca
Dünker, Nicole
Busch, Maike Anna
author_facet Doege, Annika
Steens, Rebecca
Dünker, Nicole
Busch, Maike Anna
author_sort Doege, Annika
collection PubMed
description Retinoblastoma (RB) is a primary intraocular malignancy in childhood. Relapses may develop and cause secondary cancers during later development. This study was set up to identify optimal cell culture conditions for RB cell growth in vitro and to optimize tumor growth in an in vivo model. RB cell lines (Y79 and WERI-Rb1) were cultivated under three different in vitro conditions and apoptosis, proliferation and cell growth, as well as expression profiles of two epithelial-mesenchymal transition (EMT) markers, were analyzed. EMT gene expression profiles were not generally changed, whereas apoptosis levels, tumor cell proliferation, and in vitro growth were significantly influenced by different cell culture conditions. In order to optimize the time-limited chick chorioallantoic membrane (CAM) assay, we investigated two different time points of tumor cell inoculation (embryonic development day EDD8 and EDD10) as well as three different cell concentrations. We showed that inoculation at EDD8 led to decreased tumor formation and chicken viability, whereas different cell concentrations did not change size and weight of developing tumors. Our findings demonstrate that medium conditions in vitro as well as the starting point for CAM inoculation in ovo significantly influence the experimental outcome of investigations using RB cell lines.
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spelling pubmed-89388142022-03-23 Retinoblastoma Cell Growth In Vitro and Tumor Formation In Ovo—Influence of Different Culture Conditions Doege, Annika Steens, Rebecca Dünker, Nicole Busch, Maike Anna Methods Protoc Article Retinoblastoma (RB) is a primary intraocular malignancy in childhood. Relapses may develop and cause secondary cancers during later development. This study was set up to identify optimal cell culture conditions for RB cell growth in vitro and to optimize tumor growth in an in vivo model. RB cell lines (Y79 and WERI-Rb1) were cultivated under three different in vitro conditions and apoptosis, proliferation and cell growth, as well as expression profiles of two epithelial-mesenchymal transition (EMT) markers, were analyzed. EMT gene expression profiles were not generally changed, whereas apoptosis levels, tumor cell proliferation, and in vitro growth were significantly influenced by different cell culture conditions. In order to optimize the time-limited chick chorioallantoic membrane (CAM) assay, we investigated two different time points of tumor cell inoculation (embryonic development day EDD8 and EDD10) as well as three different cell concentrations. We showed that inoculation at EDD8 led to decreased tumor formation and chicken viability, whereas different cell concentrations did not change size and weight of developing tumors. Our findings demonstrate that medium conditions in vitro as well as the starting point for CAM inoculation in ovo significantly influence the experimental outcome of investigations using RB cell lines. MDPI 2022-03-02 /pmc/articles/PMC8938814/ /pubmed/35314658 http://dx.doi.org/10.3390/mps5020021 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Doege, Annika
Steens, Rebecca
Dünker, Nicole
Busch, Maike Anna
Retinoblastoma Cell Growth In Vitro and Tumor Formation In Ovo—Influence of Different Culture Conditions
title Retinoblastoma Cell Growth In Vitro and Tumor Formation In Ovo—Influence of Different Culture Conditions
title_full Retinoblastoma Cell Growth In Vitro and Tumor Formation In Ovo—Influence of Different Culture Conditions
title_fullStr Retinoblastoma Cell Growth In Vitro and Tumor Formation In Ovo—Influence of Different Culture Conditions
title_full_unstemmed Retinoblastoma Cell Growth In Vitro and Tumor Formation In Ovo—Influence of Different Culture Conditions
title_short Retinoblastoma Cell Growth In Vitro and Tumor Formation In Ovo—Influence of Different Culture Conditions
title_sort retinoblastoma cell growth in vitro and tumor formation in ovo—influence of different culture conditions
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8938814/
https://www.ncbi.nlm.nih.gov/pubmed/35314658
http://dx.doi.org/10.3390/mps5020021
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