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In vitro Analysis of Neurospheres Derived from Glioblastoma Primary Culture: A Novel Methodology Paradigm

Glioblastomas are the most lethal primary brain tumor that frequently relapse or progress as focal masses after radiation, suggesting that a fraction of tumor cells are responsible for the tumor regrowth. The identification of a brain tumor cell subpopulation with potent tumorigenic activity support...

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Autores principales: Pavon, Lorena Favaro, Marti, Luciana C., Sibov, Tatiana Tais, Malheiros, Suzana M. F., Brandt, Reynaldo Andre, Cavalheiro, Sergio, Gamarra, Lionel F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3883037/
https://www.ncbi.nlm.nih.gov/pubmed/24432012
http://dx.doi.org/10.3389/fneur.2013.00214
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author Pavon, Lorena Favaro
Marti, Luciana C.
Sibov, Tatiana Tais
Malheiros, Suzana M. F.
Brandt, Reynaldo Andre
Cavalheiro, Sergio
Gamarra, Lionel F.
author_facet Pavon, Lorena Favaro
Marti, Luciana C.
Sibov, Tatiana Tais
Malheiros, Suzana M. F.
Brandt, Reynaldo Andre
Cavalheiro, Sergio
Gamarra, Lionel F.
author_sort Pavon, Lorena Favaro
collection PubMed
description Glioblastomas are the most lethal primary brain tumor that frequently relapse or progress as focal masses after radiation, suggesting that a fraction of tumor cells are responsible for the tumor regrowth. The identification of a brain tumor cell subpopulation with potent tumorigenic activity supports the cancer stem cell hypothesis in solid tumors. The goal of this study is to determine a methodology for the establishment of primary human glioblastoma cell lines. Our aim is achieved by taking the following approaches: (i) the establishment of primary glioblastoma cell culture; (ii) isolation of neurospheres derived from glioblastoma primary cultures; (iii) selection of CD133 cells from neurospheres, (iv) formation of subspheres in the CD133-positive population, (v) study of the expression level of GFAP, CD133, Nestin, Nanog, CD34, Sox2, CD44, and CD90 markers on tumor subspheres. Hence, we described a successful method for isolation of CD133-positive cell population and establishment of glioblastoma neurospheres from this primary culture, which are more robust than the ones derived straight from the tumor. Pointed out that the neurospheres derived from glioblastoma primary culture showed 29% more cells expressing CD133 then the ones straight tumor-derived, denoting a higher concentration of CD133-positive cells in the neurospheres derived from glioblastoma primary culture. These CD133-positive fractions were able to further generate subspheres. The subspheres derived from glioblastoma primary culture presented a well-defined morphology while the ones derived from the fresh tumor were sparce and less robust. And the negative fraction of CD133 cells was unable to generate subspheres. The tumor subspheres expressed GFAP, CD133, Nestin, Nanog, CD44, and CD90. Also, the present study describes an optimization of neurospheres/subspheres isolation from glioblastoma primary culture by selection of CD133-positive adherent stem cell.
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spelling pubmed-38830372014-01-15 In vitro Analysis of Neurospheres Derived from Glioblastoma Primary Culture: A Novel Methodology Paradigm Pavon, Lorena Favaro Marti, Luciana C. Sibov, Tatiana Tais Malheiros, Suzana M. F. Brandt, Reynaldo Andre Cavalheiro, Sergio Gamarra, Lionel F. Front Neurol Neuroscience Glioblastomas are the most lethal primary brain tumor that frequently relapse or progress as focal masses after radiation, suggesting that a fraction of tumor cells are responsible for the tumor regrowth. The identification of a brain tumor cell subpopulation with potent tumorigenic activity supports the cancer stem cell hypothesis in solid tumors. The goal of this study is to determine a methodology for the establishment of primary human glioblastoma cell lines. Our aim is achieved by taking the following approaches: (i) the establishment of primary glioblastoma cell culture; (ii) isolation of neurospheres derived from glioblastoma primary cultures; (iii) selection of CD133 cells from neurospheres, (iv) formation of subspheres in the CD133-positive population, (v) study of the expression level of GFAP, CD133, Nestin, Nanog, CD34, Sox2, CD44, and CD90 markers on tumor subspheres. Hence, we described a successful method for isolation of CD133-positive cell population and establishment of glioblastoma neurospheres from this primary culture, which are more robust than the ones derived straight from the tumor. Pointed out that the neurospheres derived from glioblastoma primary culture showed 29% more cells expressing CD133 then the ones straight tumor-derived, denoting a higher concentration of CD133-positive cells in the neurospheres derived from glioblastoma primary culture. These CD133-positive fractions were able to further generate subspheres. The subspheres derived from glioblastoma primary culture presented a well-defined morphology while the ones derived from the fresh tumor were sparce and less robust. And the negative fraction of CD133 cells was unable to generate subspheres. The tumor subspheres expressed GFAP, CD133, Nestin, Nanog, CD44, and CD90. Also, the present study describes an optimization of neurospheres/subspheres isolation from glioblastoma primary culture by selection of CD133-positive adherent stem cell. Frontiers Media S.A. 2014-01-07 /pmc/articles/PMC3883037/ /pubmed/24432012 http://dx.doi.org/10.3389/fneur.2013.00214 Text en Copyright © 2014 Pavon, Marti, Sibov, Malheiros, Brandt, Cavalheiro and Gamarra. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Neuroscience
Pavon, Lorena Favaro
Marti, Luciana C.
Sibov, Tatiana Tais
Malheiros, Suzana M. F.
Brandt, Reynaldo Andre
Cavalheiro, Sergio
Gamarra, Lionel F.
In vitro Analysis of Neurospheres Derived from Glioblastoma Primary Culture: A Novel Methodology Paradigm
title In vitro Analysis of Neurospheres Derived from Glioblastoma Primary Culture: A Novel Methodology Paradigm
title_full In vitro Analysis of Neurospheres Derived from Glioblastoma Primary Culture: A Novel Methodology Paradigm
title_fullStr In vitro Analysis of Neurospheres Derived from Glioblastoma Primary Culture: A Novel Methodology Paradigm
title_full_unstemmed In vitro Analysis of Neurospheres Derived from Glioblastoma Primary Culture: A Novel Methodology Paradigm
title_short In vitro Analysis of Neurospheres Derived from Glioblastoma Primary Culture: A Novel Methodology Paradigm
title_sort in vitro analysis of neurospheres derived from glioblastoma primary culture: a novel methodology paradigm
topic Neuroscience
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3883037/
https://www.ncbi.nlm.nih.gov/pubmed/24432012
http://dx.doi.org/10.3389/fneur.2013.00214
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