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Cell fusion in tumor progression: the isolation of cell fusion products by physical methods

BACKGROUND: Cell fusion induced by polyethylene glycol (PEG) is an efficient but poorly controlled procedure for obtaining somatic cell hybrids used in gene mapping, monoclonal antibody production, and tumour immunotherapy. Genetic selection techniques and fluorescent cell sorting are usually employ...

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Detalles Bibliográficos
Autores principales: Pedrazzoli, Filippo, Chrysantzas, Iraklis, Dezzani, Luca, Rosti, Vittorio, Vincitorio, Massimo, Sitar, Giammaria
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3187729/
https://www.ncbi.nlm.nih.gov/pubmed/21933375
http://dx.doi.org/10.1186/1475-2867-11-32
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author Pedrazzoli, Filippo
Chrysantzas, Iraklis
Dezzani, Luca
Rosti, Vittorio
Vincitorio, Massimo
Sitar, Giammaria
author_facet Pedrazzoli, Filippo
Chrysantzas, Iraklis
Dezzani, Luca
Rosti, Vittorio
Vincitorio, Massimo
Sitar, Giammaria
author_sort Pedrazzoli, Filippo
collection PubMed
description BACKGROUND: Cell fusion induced by polyethylene glycol (PEG) is an efficient but poorly controlled procedure for obtaining somatic cell hybrids used in gene mapping, monoclonal antibody production, and tumour immunotherapy. Genetic selection techniques and fluorescent cell sorting are usually employed to isolate cell fusion products, but both procedures have several drawbacks. RESULTS: Here we describe a simple improvement in PEG-mediated cell fusion that was obtained by modifying the standard single-step procedure. We found that the use of two PEG undertreatments obtains a better yield of cell fusion products than the standard method, and most of these products are bi- or trinucleated polykaryocytes. Fusion rate was quantified using fluorescent cell staining microscopy. We used this improved cell fusion and cell isolation method to compare giant cells obtained in vitro and giant cells obtained in vivo from patients with Hodgkin's disease and erythroleukemia. CONCLUSIONS: In the present study we show how to improve PEG-mediated cell fusion and that cell separation by velocity sedimentation offers a simple alternative for the efficient purification of cell fusion products and to investigate giant cell formation in tumor development.
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spelling pubmed-31877292011-10-06 Cell fusion in tumor progression: the isolation of cell fusion products by physical methods Pedrazzoli, Filippo Chrysantzas, Iraklis Dezzani, Luca Rosti, Vittorio Vincitorio, Massimo Sitar, Giammaria Cancer Cell Int Primary Research BACKGROUND: Cell fusion induced by polyethylene glycol (PEG) is an efficient but poorly controlled procedure for obtaining somatic cell hybrids used in gene mapping, monoclonal antibody production, and tumour immunotherapy. Genetic selection techniques and fluorescent cell sorting are usually employed to isolate cell fusion products, but both procedures have several drawbacks. RESULTS: Here we describe a simple improvement in PEG-mediated cell fusion that was obtained by modifying the standard single-step procedure. We found that the use of two PEG undertreatments obtains a better yield of cell fusion products than the standard method, and most of these products are bi- or trinucleated polykaryocytes. Fusion rate was quantified using fluorescent cell staining microscopy. We used this improved cell fusion and cell isolation method to compare giant cells obtained in vitro and giant cells obtained in vivo from patients with Hodgkin's disease and erythroleukemia. CONCLUSIONS: In the present study we show how to improve PEG-mediated cell fusion and that cell separation by velocity sedimentation offers a simple alternative for the efficient purification of cell fusion products and to investigate giant cell formation in tumor development. BioMed Central 2011-09-20 /pmc/articles/PMC3187729/ /pubmed/21933375 http://dx.doi.org/10.1186/1475-2867-11-32 Text en Copyright ©2011 Pedrazzoli et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Primary Research
Pedrazzoli, Filippo
Chrysantzas, Iraklis
Dezzani, Luca
Rosti, Vittorio
Vincitorio, Massimo
Sitar, Giammaria
Cell fusion in tumor progression: the isolation of cell fusion products by physical methods
title Cell fusion in tumor progression: the isolation of cell fusion products by physical methods
title_full Cell fusion in tumor progression: the isolation of cell fusion products by physical methods
title_fullStr Cell fusion in tumor progression: the isolation of cell fusion products by physical methods
title_full_unstemmed Cell fusion in tumor progression: the isolation of cell fusion products by physical methods
title_short Cell fusion in tumor progression: the isolation of cell fusion products by physical methods
title_sort cell fusion in tumor progression: the isolation of cell fusion products by physical methods
topic Primary Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3187729/
https://www.ncbi.nlm.nih.gov/pubmed/21933375
http://dx.doi.org/10.1186/1475-2867-11-32
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