Immunocytochemistry Analysis of HepG2 Cell 3D Culture Encapsulated as Spheroids in Alginate Beads
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Outros Autores: | , , |
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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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 |