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

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dc.contributor.author Skarkova, Veronika cze
dc.contributor.author Krupova, Marketa cze
dc.contributor.author Vitovcova, Barbora cze
dc.contributor.author Skarka, Adam cze
dc.contributor.author Kasparova, Petra cze
dc.contributor.author Krupa, Petr cze
dc.contributor.author Kralova, Vera cze
dc.contributor.author Rudolf, Emil cze
dc.date.accessioned 2019-11-21T08:29:54Z
dc.date.available 2019-11-21T08:29:54Z
dc.date.issued 2019 eng
dc.identifier.issn 1422-0067 eng
dc.identifier.uri http://hdl.handle.net/20.500.12603/67
dc.description.abstract 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 eng
dc.format p. "Article Number: 4630" eng
dc.language.iso eng eng
dc.publisher MDPI eng
dc.relation.ispartof INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, volume 20, issue: 18 eng
dc.rights Práce není přístupná eng
dc.subject glioblastoma multiforme eng
dc.subject cell isolation eng
dc.subject temozolomide eng
dc.subject resistance eng
dc.subject glioblastoma multiforme cze
dc.subject izolace buněk cze
dc.subject temozolomid cze
dc.subject rezistence cze
dc.title The Evaluation of Glioblastoma Cell Dissociation and Its Influence on Its Behavior eng
dc.title.alternative Hodnocení disociace buněk glioblastomu a jeho vliv na jejich chování cze
dc.type article eng
dc.identifier.obd 43875655 eng
dc.identifier.wos 000489100500309 eng
dc.identifier.doi 10.3390/ijms20184630 eng
dc.description.abstract-translated 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 cze
dc.publicationstatus postprint eng
dc.peerreviewed yes eng


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