Efficient isolation and proliferation of human adipose-derived mesenchymal stromal cells in xeno-free conditions

Detalhes bibliográficos
Autor(a) principal: Fuoco, Natalia Langenfeld
Data de Publicação: 2020
Outros Autores: de Oliveira, Rafael Guilen, Marcelino, Monica Yonashiro, Stessuk, Talita, Sakalem, Marna Eliana [UNESP], Medina, Denis Aloisio Lopes, Modotti, Waldir Pereira, Forte, Andresa, Ribeiro-Paes, João Tadeu [UNESP]
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1007/s11033-020-05322-9
http://hdl.handle.net/11449/198629
Resumo: Classical methods used for culture of adipose-derived mesenchymal stromal cells (ADSCs) use xenobiotic components, which may present a potential risk for biological contamination and/or elicit immunological reactions. Therefore, the aim of this study was to establish a xeno-free methodology for the isolation and proliferation of human ADSCs (hADSCs). hADSCs were isolated by enzymatic digestion or mechanical dissociation and cultured in the presence of fetal bovine serum or human platelet lysate. Proliferation curves were performed as a function of time from the cell culture and used to calculate the population doubling time. Immunophenotyping and differentiation tests were used to identify and characterize the hADSCs. Human ADSCs isolated and cultured in conventional or xenobiotic-free conditions peaked at different days but achieved similar maximum proliferation. The hADSCs differentiation ability was similar in all groups. The characterization of hADSCs by flow cytometry showed low contamination of the cultures by other cell types. The xenobiotic-free methodology described in this study is a feasible and reproducible alternative for isolation and proliferation of hADSCs. This methodology is in accordance with the recommendations of the National Health Surveillance Agency, which proposes avoidance of xenobiotic products.
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spelling Efficient isolation and proliferation of human adipose-derived mesenchymal stromal cells in xeno-free conditionsAdipose tissueFetal bovine serum (FBS)Mesenchymal stromal cellsPlatelet lysateStem cellsClassical methods used for culture of adipose-derived mesenchymal stromal cells (ADSCs) use xenobiotic components, which may present a potential risk for biological contamination and/or elicit immunological reactions. Therefore, the aim of this study was to establish a xeno-free methodology for the isolation and proliferation of human ADSCs (hADSCs). hADSCs were isolated by enzymatic digestion or mechanical dissociation and cultured in the presence of fetal bovine serum or human platelet lysate. Proliferation curves were performed as a function of time from the cell culture and used to calculate the population doubling time. Immunophenotyping and differentiation tests were used to identify and characterize the hADSCs. Human ADSCs isolated and cultured in conventional or xenobiotic-free conditions peaked at different days but achieved similar maximum proliferation. The hADSCs differentiation ability was similar in all groups. The characterization of hADSCs by flow cytometry showed low contamination of the cultures by other cell types. The xenobiotic-free methodology described in this study is a feasible and reproducible alternative for isolation and proliferation of hADSCs. This methodology is in accordance with the recommendations of the National Health Surveillance Agency, which proposes avoidance of xenobiotic products.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Biotechnology Interunits Post-Graduation Program Biomedical Science Institute University of São Paulo (USP)Institute of Medical-Hospital Service (IAM)São Lucas – Cell Therapy GroupGenetics and Cell Therapy Laboratory (GenTe Cel) São Paulo State University (Unesp)Laboratório de Genética e Terapia Celular – GenTe Cel Departamento de Biotecnologia – Unesp, Av. Dom Antonio, 2100Genetics and Cell Therapy Laboratory (GenTe Cel) São Paulo State University (Unesp)Laboratório de Genética e Terapia Celular – GenTe Cel Departamento de Biotecnologia – Unesp, Av. Dom Antonio, 2100Universidade de São Paulo (USP)Institute of Medical-Hospital Service (IAM)São Lucas – Cell Therapy GroupUniversidade Estadual Paulista (Unesp)Fuoco, Natalia Langenfeldde Oliveira, Rafael GuilenMarcelino, Monica YonashiroStessuk, TalitaSakalem, Marna Eliana [UNESP]Medina, Denis Aloisio LopesModotti, Waldir PereiraForte, AndresaRibeiro-Paes, João Tadeu [UNESP]2020-12-12T01:18:03Z2020-12-12T01:18:03Z2020-04-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article2475-2486http://dx.doi.org/10.1007/s11033-020-05322-9Molecular Biology Reports, v. 47, n. 4, p. 2475-2486, 2020.1573-49780301-4851http://hdl.handle.net/11449/19862910.1007/s11033-020-05322-92-s2.0-85081588847Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengMolecular Biology Reportsinfo:eu-repo/semantics/openAccess2021-10-22T17:50:50Zoai:repositorio.unesp.br:11449/198629Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T20:17:04.070084Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Efficient isolation and proliferation of human adipose-derived mesenchymal stromal cells in xeno-free conditions
title Efficient isolation and proliferation of human adipose-derived mesenchymal stromal cells in xeno-free conditions
spellingShingle Efficient isolation and proliferation of human adipose-derived mesenchymal stromal cells in xeno-free conditions
Fuoco, Natalia Langenfeld
Adipose tissue
Fetal bovine serum (FBS)
Mesenchymal stromal cells
Platelet lysate
Stem cells
title_short Efficient isolation and proliferation of human adipose-derived mesenchymal stromal cells in xeno-free conditions
title_full Efficient isolation and proliferation of human adipose-derived mesenchymal stromal cells in xeno-free conditions
title_fullStr Efficient isolation and proliferation of human adipose-derived mesenchymal stromal cells in xeno-free conditions
title_full_unstemmed Efficient isolation and proliferation of human adipose-derived mesenchymal stromal cells in xeno-free conditions
title_sort Efficient isolation and proliferation of human adipose-derived mesenchymal stromal cells in xeno-free conditions
author Fuoco, Natalia Langenfeld
author_facet Fuoco, Natalia Langenfeld
de Oliveira, Rafael Guilen
Marcelino, Monica Yonashiro
Stessuk, Talita
Sakalem, Marna Eliana [UNESP]
Medina, Denis Aloisio Lopes
Modotti, Waldir Pereira
Forte, Andresa
Ribeiro-Paes, João Tadeu [UNESP]
author_role author
author2 de Oliveira, Rafael Guilen
Marcelino, Monica Yonashiro
Stessuk, Talita
Sakalem, Marna Eliana [UNESP]
Medina, Denis Aloisio Lopes
Modotti, Waldir Pereira
Forte, Andresa
Ribeiro-Paes, João Tadeu [UNESP]
author2_role author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade de São Paulo (USP)
Institute of Medical-Hospital Service (IAM)
São Lucas – Cell Therapy Group
Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Fuoco, Natalia Langenfeld
de Oliveira, Rafael Guilen
Marcelino, Monica Yonashiro
Stessuk, Talita
Sakalem, Marna Eliana [UNESP]
Medina, Denis Aloisio Lopes
Modotti, Waldir Pereira
Forte, Andresa
Ribeiro-Paes, João Tadeu [UNESP]
dc.subject.por.fl_str_mv Adipose tissue
Fetal bovine serum (FBS)
Mesenchymal stromal cells
Platelet lysate
Stem cells
topic Adipose tissue
Fetal bovine serum (FBS)
Mesenchymal stromal cells
Platelet lysate
Stem cells
description Classical methods used for culture of adipose-derived mesenchymal stromal cells (ADSCs) use xenobiotic components, which may present a potential risk for biological contamination and/or elicit immunological reactions. Therefore, the aim of this study was to establish a xeno-free methodology for the isolation and proliferation of human ADSCs (hADSCs). hADSCs were isolated by enzymatic digestion or mechanical dissociation and cultured in the presence of fetal bovine serum or human platelet lysate. Proliferation curves were performed as a function of time from the cell culture and used to calculate the population doubling time. Immunophenotyping and differentiation tests were used to identify and characterize the hADSCs. Human ADSCs isolated and cultured in conventional or xenobiotic-free conditions peaked at different days but achieved similar maximum proliferation. The hADSCs differentiation ability was similar in all groups. The characterization of hADSCs by flow cytometry showed low contamination of the cultures by other cell types. The xenobiotic-free methodology described in this study is a feasible and reproducible alternative for isolation and proliferation of hADSCs. This methodology is in accordance with the recommendations of the National Health Surveillance Agency, which proposes avoidance of xenobiotic products.
publishDate 2020
dc.date.none.fl_str_mv 2020-12-12T01:18:03Z
2020-12-12T01:18:03Z
2020-04-01
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1007/s11033-020-05322-9
Molecular Biology Reports, v. 47, n. 4, p. 2475-2486, 2020.
1573-4978
0301-4851
http://hdl.handle.net/11449/198629
10.1007/s11033-020-05322-9
2-s2.0-85081588847
url http://dx.doi.org/10.1007/s11033-020-05322-9
http://hdl.handle.net/11449/198629
identifier_str_mv Molecular Biology Reports, v. 47, n. 4, p. 2475-2486, 2020.
1573-4978
0301-4851
10.1007/s11033-020-05322-9
2-s2.0-85081588847
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Molecular Biology Reports
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 2475-2486
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
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