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The Evaluation of Glioblastoma Cell Dissociation and Its Influence on Its Behavior

Purpose: Primary cell lines are a valuable tool for evaluation of tumor behavior or sensitivity to anticancer treatment and appropriate dissociation of cells could preserve genomic profile of the original tissue. The main aim of our study was to compare the influence of two methods of glioblastoma m...

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Autores principales: Skarkova, Veronika, Krupova, Marketa, Vitovcova, Barbora, Skarka, Adam, Kasparova, Petra, Krupa, Petr, Kralova, Vera, Rudolf, Emil
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6770747/
https://www.ncbi.nlm.nih.gov/pubmed/31540507
http://dx.doi.org/10.3390/ijms20184630
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author Skarkova, Veronika
Krupova, Marketa
Vitovcova, Barbora
Skarka, Adam
Kasparova, Petra
Krupa, Petr
Kralova, Vera
Rudolf, Emil
author_facet Skarkova, Veronika
Krupova, Marketa
Vitovcova, Barbora
Skarka, Adam
Kasparova, Petra
Krupa, Petr
Kralova, Vera
Rudolf, Emil
author_sort Skarkova, Veronika
collection PubMed
description Purpose: Primary cell lines are a valuable tool for evaluation of tumor behavior or sensitivity to anticancer treatment and appropriate dissociation of cells could preserve genomic profile of the original tissue. The main aim of our study was to compare the influence of two methods of glioblastoma multiforme (GBM) cell derivation (mechanic—MD; enzymatic—ED) on basic biological properties of thus derived cells and correlate them to the ones obtained from stabilized GBM cell line A-172. Methods: Cell proliferation and migration (xCELLigence Real-Time Cell Analysis), expression of microRNAs and protein markers (RT-PCR and Western blotting), morphology (phase contrast and fluorescent microscopy), and accumulation of temozolomide (TMZ) and its metabolite 5-aminoimidazole-4-carboxamide (AIC) inside the cells (LC-MS analysis) were carried out in five different samples of GBM (GBM1, GBM2, GBM32, GBM33, GBM34), with each of them processed by MD and ED types of isolations. The same analyses were done in the A-172 cell line too. Results: Primary GBM cells obtained by ED or MD approaches significantly differ in biological behavior and properties of these cells. Unlike in primary MD GBM cells, higher proliferation, as well as migration, was observed in primary ED GBM cells, which were also associated with the acquired mesenchymal phenotype and higher sensitivity to TMZ. Finally, the same analyses of stabilized GBM cell line A-172 revealed several important differences in measured parameters. Conclusions: GBM cells obtained by MD and ED dissociation show considerable heterogeneity, but based on our results, MD approach should be the preferred method of primary GBM cell isolation
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spelling pubmed-67707472019-10-30 The Evaluation of Glioblastoma Cell Dissociation and Its Influence on Its Behavior Skarkova, Veronika Krupova, Marketa Vitovcova, Barbora Skarka, Adam Kasparova, Petra Krupa, Petr Kralova, Vera Rudolf, Emil Int J Mol Sci Article Purpose: Primary cell lines are a valuable tool for evaluation of tumor behavior or sensitivity to anticancer treatment and appropriate dissociation of cells could preserve genomic profile of the original tissue. The main aim of our study was to compare the influence of two methods of glioblastoma multiforme (GBM) cell derivation (mechanic—MD; enzymatic—ED) on basic biological properties of thus derived cells and correlate them to the ones obtained from stabilized GBM cell line A-172. Methods: Cell proliferation and migration (xCELLigence Real-Time Cell Analysis), expression of microRNAs and protein markers (RT-PCR and Western blotting), morphology (phase contrast and fluorescent microscopy), and accumulation of temozolomide (TMZ) and its metabolite 5-aminoimidazole-4-carboxamide (AIC) inside the cells (LC-MS analysis) were carried out in five different samples of GBM (GBM1, GBM2, GBM32, GBM33, GBM34), with each of them processed by MD and ED types of isolations. The same analyses were done in the A-172 cell line too. Results: Primary GBM cells obtained by ED or MD approaches significantly differ in biological behavior and properties of these cells. Unlike in primary MD GBM cells, higher proliferation, as well as migration, was observed in primary ED GBM cells, which were also associated with the acquired mesenchymal phenotype and higher sensitivity to TMZ. Finally, the same analyses of stabilized GBM cell line A-172 revealed several important differences in measured parameters. Conclusions: GBM cells obtained by MD and ED dissociation show considerable heterogeneity, but based on our results, MD approach should be the preferred method of primary GBM cell isolation MDPI 2019-09-18 /pmc/articles/PMC6770747/ /pubmed/31540507 http://dx.doi.org/10.3390/ijms20184630 Text en © 2019 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Skarkova, Veronika
Krupova, Marketa
Vitovcova, Barbora
Skarka, Adam
Kasparova, Petra
Krupa, Petr
Kralova, Vera
Rudolf, Emil
The Evaluation of Glioblastoma Cell Dissociation and Its Influence on Its Behavior
title The Evaluation of Glioblastoma Cell Dissociation and Its Influence on Its Behavior
title_full The Evaluation of Glioblastoma Cell Dissociation and Its Influence on Its Behavior
title_fullStr The Evaluation of Glioblastoma Cell Dissociation and Its Influence on Its Behavior
title_full_unstemmed The Evaluation of Glioblastoma Cell Dissociation and Its Influence on Its Behavior
title_short The Evaluation of Glioblastoma Cell Dissociation and Its Influence on Its Behavior
title_sort evaluation of glioblastoma cell dissociation and its influence on its behavior
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6770747/
https://www.ncbi.nlm.nih.gov/pubmed/31540507
http://dx.doi.org/10.3390/ijms20184630
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