Efficient isolation and proliferation of human adipose-derived mesenchymal stromal cells in xeno-free conditions
| Autor: | Marna Eliana Sakalem, Andresa Forte, Monica Yonashiro Marcelino, Waldir Pereira Modotti, Denis Aloisio Lopes Medina, João Tadeu Ribeiro-Paes, Talita Stessuk, Rafael Guilen de Oliveira, Natalia Langenfeld Fuoco |
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| Přispěvatelé: | Universidade de São Paulo (USP), Institute of Medical-Hospital Service (IAM), São Lucas – Cell Therapy Group, Universidade Estadual Paulista (Unesp) |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Cell Culture Techniques Mesenchymal stromal cells Adipose tissue Stem cells Flow cytometry Immunophenotyping Xenobiotics 03 medical and health sciences 0302 clinical medicine Genetics medicine Doubling time Humans Platelet lysate Molecular Biology Cells Cultured Cell Proliferation medicine.diagnostic_test Chemistry Mesenchymal stem cell Cell Differentiation Mesenchymal Stem Cells General Medicine Flow Cytometry Cell biology Culture Media 030104 developmental biology Fetal bovine serum (FBS) Adipose Tissue Cell culture 030220 oncology & carcinogenesis Stem cell Fetal bovine serum |
| Zdroj: | Scopus Repositório Institucional da UNESP Universidade Estadual Paulista (UNESP) instacron:UNESP |
| Popis: | Made available in DSpace on 2020-12-12T01:18:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-04-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) 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. Biotechnology Interunits Post-Graduation Program Biomedical Science Institute University of São Paulo (USP) Institute of Medical-Hospital Service (IAM) São Lucas – Cell Therapy Group Genetics 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, 2100 Genetics 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, 2100 |
| Databáze: | OpenAIRE |
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