Efficient isolation and proliferation of human adipose-derived mesenchymal stromal cells in xeno-free conditions
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , , , , , , |
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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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 |
|
_version_ |
1808129183381979136 |