Immunocytochemistry Analysis of HepG2 Cell 3D Culture Encapsulated as Spheroids in Alginate Beads

Detalhes bibliográficos
Autor(a) principal: Miranda, Raul Ghiraldelli
Data de Publicação: 2021
Outros Autores: Ferraz, Elisa Raquel Anastácio, Pereira, Lilian Cristina [UNESP], Dorta, Daniel Junqueira [UNESP]
Tipo de documento: Capítulo de livro
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1007/978-1-0716-1091-6_14
http://hdl.handle.net/11449/207167
Resumo: 3D Cell culture is an alternative to animal use in many drug development and toxicity studies. The 3D cell culture can mimic and reproduce the original tissue microenvironment, morphology, and mechanical and physiological characteristics, to provide a more realistic and reliable response as compared to two-dimensional cultures. 3D cell culture encapsulated in alginate beads is a very simple and relatively inexpensive tool that is easy to handle and to maintain. The alginate beads function as a scaffold that imprisons cells and allows 3D cell growth, to generate spheroids that can have greater genic expression and cell–cell communication as a nano or microtissue. The HepG2 cell line is a human hepatocellular carcinoma cell derivative. HepG2 cells preserve several of the characteristics of hepatocytes and are therefore often used in toxicity studies. Here, we describe HepG2 cell encapsulation in alginate beads and analyze the resulting spheroids formed within the alginate beads by immunocytochemistry, by staining a certain structure with a specific antibody coupled with a fluorophore. This method preserves the beads and enables cell analysis by confocal microscopy.
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spelling Immunocytochemistry Analysis of HepG2 Cell 3D Culture Encapsulated as Spheroids in Alginate Beads3D cultureAlginate beadsConfocal microscopyEncapsulationHepG2Immunocytochemistry3D Cell culture is an alternative to animal use in many drug development and toxicity studies. The 3D cell culture can mimic and reproduce the original tissue microenvironment, morphology, and mechanical and physiological characteristics, to provide a more realistic and reliable response as compared to two-dimensional cultures. 3D cell culture encapsulated in alginate beads is a very simple and relatively inexpensive tool that is easy to handle and to maintain. The alginate beads function as a scaffold that imprisons cells and allows 3D cell growth, to generate spheroids that can have greater genic expression and cell–cell communication as a nano or microtissue. The HepG2 cell line is a human hepatocellular carcinoma cell derivative. HepG2 cells preserve several of the characteristics of hepatocytes and are therefore often used in toxicity studies. Here, we describe HepG2 cell encapsulation in alginate beads and analyze the resulting spheroids formed within the alginate beads by immunocytochemistry, by staining a certain structure with a specific antibody coupled with a fluorophore. This method preserves the beads and enables cell analysis by confocal microscopy.Departamento de Química Faculdade de Filosofia Ciências e Letras de Ribeirão Preto Universidade de São PauloDepartment of Clinical Toxicological and Bromatological Analysis Faculty of Pharmaceutical Sciences of Ribeirão Preto University of São PauloFaculdade de Farmácia Universidade Federal FluminenseDepartment of Bioprocesses and Biotechnology Faculty of Agronomic Sciences of Botucatu São Paulo State UniversityCenter for Evaluation of Environmental Impact on Human Health (TOXICAM) BotucatuFFCLRP-USP Department of Chemistry Faculty of Philosophy Sciences and Letters of Ribeirão Preto University of São Paulo Ribeirão PretoInstituto Nacional de Tecnologias Alternativas de Detecção Avaliação Toxicologicae Remoção de Micropututantes e Radioativos (INCT-DATREM) Unesp Instituto de QuímicaDepartment of Bioprocesses and Biotechnology Faculty of Agronomic Sciences of Botucatu São Paulo State UniversityInstituto Nacional de Tecnologias Alternativas de Detecção Avaliação Toxicologicae Remoção de Micropututantes e Radioativos (INCT-DATREM) Unesp Instituto de QuímicaUniversidade de São Paulo (USP)Universidade Federal Fluminense (UFF)Universidade Estadual Paulista (Unesp)BotucatuMiranda, Raul GhiraldelliFerraz, Elisa Raquel AnastácioPereira, Lilian Cristina [UNESP]Dorta, Daniel Junqueira [UNESP]2021-06-25T10:50:04Z2021-06-25T10:50:04Z2021-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/bookPart197-206http://dx.doi.org/10.1007/978-1-0716-1091-6_14Methods in Molecular Biology, v. 2240, p. 197-206.1940-60291064-3745http://hdl.handle.net/11449/20716710.1007/978-1-0716-1091-6_142-s2.0-85099708839Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengMethods in Molecular Biologyinfo:eu-repo/semantics/openAccess2021-10-23T16:22:43Zoai:repositorio.unesp.br:11449/207167Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T13:53:19.047439Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Immunocytochemistry Analysis of HepG2 Cell 3D Culture Encapsulated as Spheroids in Alginate Beads
title Immunocytochemistry Analysis of HepG2 Cell 3D Culture Encapsulated as Spheroids in Alginate Beads
spellingShingle Immunocytochemistry Analysis of HepG2 Cell 3D Culture Encapsulated as Spheroids in Alginate Beads
Miranda, Raul Ghiraldelli
3D culture
Alginate beads
Confocal microscopy
Encapsulation
HepG2
Immunocytochemistry
title_short Immunocytochemistry Analysis of HepG2 Cell 3D Culture Encapsulated as Spheroids in Alginate Beads
title_full Immunocytochemistry Analysis of HepG2 Cell 3D Culture Encapsulated as Spheroids in Alginate Beads
title_fullStr Immunocytochemistry Analysis of HepG2 Cell 3D Culture Encapsulated as Spheroids in Alginate Beads
title_full_unstemmed Immunocytochemistry Analysis of HepG2 Cell 3D Culture Encapsulated as Spheroids in Alginate Beads
title_sort Immunocytochemistry Analysis of HepG2 Cell 3D Culture Encapsulated as Spheroids in Alginate Beads
author Miranda, Raul Ghiraldelli
author_facet Miranda, Raul Ghiraldelli
Ferraz, Elisa Raquel Anastácio
Pereira, Lilian Cristina [UNESP]
Dorta, Daniel Junqueira [UNESP]
author_role author
author2 Ferraz, Elisa Raquel Anastácio
Pereira, Lilian Cristina [UNESP]
Dorta, Daniel Junqueira [UNESP]
author2_role author
author
author
dc.contributor.none.fl_str_mv Universidade de São Paulo (USP)
Universidade Federal Fluminense (UFF)
Universidade Estadual Paulista (Unesp)
Botucatu
dc.contributor.author.fl_str_mv Miranda, Raul Ghiraldelli
Ferraz, Elisa Raquel Anastácio
Pereira, Lilian Cristina [UNESP]
Dorta, Daniel Junqueira [UNESP]
dc.subject.por.fl_str_mv 3D culture
Alginate beads
Confocal microscopy
Encapsulation
HepG2
Immunocytochemistry
topic 3D culture
Alginate beads
Confocal microscopy
Encapsulation
HepG2
Immunocytochemistry
description 3D Cell culture is an alternative to animal use in many drug development and toxicity studies. The 3D cell culture can mimic and reproduce the original tissue microenvironment, morphology, and mechanical and physiological characteristics, to provide a more realistic and reliable response as compared to two-dimensional cultures. 3D cell culture encapsulated in alginate beads is a very simple and relatively inexpensive tool that is easy to handle and to maintain. The alginate beads function as a scaffold that imprisons cells and allows 3D cell growth, to generate spheroids that can have greater genic expression and cell–cell communication as a nano or microtissue. The HepG2 cell line is a human hepatocellular carcinoma cell derivative. HepG2 cells preserve several of the characteristics of hepatocytes and are therefore often used in toxicity studies. Here, we describe HepG2 cell encapsulation in alginate beads and analyze the resulting spheroids formed within the alginate beads by immunocytochemistry, by staining a certain structure with a specific antibody coupled with a fluorophore. This method preserves the beads and enables cell analysis by confocal microscopy.
publishDate 2021
dc.date.none.fl_str_mv 2021-06-25T10:50:04Z
2021-06-25T10:50:04Z
2021-01-01
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/bookPart
format bookPart
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1007/978-1-0716-1091-6_14
Methods in Molecular Biology, v. 2240, p. 197-206.
1940-6029
1064-3745
http://hdl.handle.net/11449/207167
10.1007/978-1-0716-1091-6_14
2-s2.0-85099708839
url http://dx.doi.org/10.1007/978-1-0716-1091-6_14
http://hdl.handle.net/11449/207167
identifier_str_mv Methods in Molecular Biology, v. 2240, p. 197-206.
1940-6029
1064-3745
10.1007/978-1-0716-1091-6_14
2-s2.0-85099708839
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Methods in Molecular Biology
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 197-206
dc.source.none.fl_str_mv Scopus
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
_version_ 1808128287214403584