Adipose-derived stem cells (ASCs) culture in spinner flask: improving the parameters of culture in a microcarrier-based system.

Autor: Simão VA; Department of Genetics, School of Medicine, University of São Paulo, Ribeirão Preto, São Paulo, Brazil. viniciusmorma@gmail.com., Brand H; Department of Biotechnology, School of Sciences and Letters, São Paulo State University (UNESP), Assis, São Paulo, Brazil., da Silveira-Antunes RN; Department of Hematology, Hemocenter, School of Medicine of Marilia, Marilia, São Paulo, Brazil., Fukasawa JT; Department of Hematology, Hemocenter, School of Medicine of Marilia, Marilia, São Paulo, Brazil., Leme J; Center for Development and Innovation, Laboratory of Viral Biotechnology, Butantan Institute, São Paulo, São Paulo, Brazil., Tonso A; Department of Chemical Engineering, Polytechnic School, University of São Paulo, São Paulo, São Paulo, Brazil., Ribeiro-Paes JT; Department of Biotechnology, School of Sciences and Letters, São Paulo State University (UNESP), Assis, São Paulo, Brazil.
Jazyk: angličtina
Zdroj: Biotechnology letters [Biotechnol Lett] 2023 Jul; Vol. 45 (7), pp. 823-846. Date of Electronic Publication: 2023 May 12.
DOI: 10.1007/s10529-023-03367-x
Abstrakt: Prior to clinical use, extensive in vitro proliferation of human adipose-derived stem cells (ASCs) is required. Among the current options, spinner-type stirred flasks, which use microcarriers to increase the yield of adherent cells, are recommended. Here, we propose a methodology for ASCs proliferation through cell suspension culture using Cultispher-S ® microcarriers (MC) under agitation in a spinner flask, with the aim of establishing a system that reconciles the efficiency of cell yield with high viability of the culture during two distinct phases: seeding and proliferation. The results showed that cell adhesion was potentiated under intermittent stirring at 70 rpm in the presence of 10% FBS for an initial cell concentration of 2.4 × 10 4 cells/mL in the initial 24 h of cultivation. In the proliferation phase, kinetic analysis showed that cell growth was higher under continuous agitation at 50 rpm with a culture medium renewal regime of 50% every 72 h, which was sufficient to maintain the culture at optimal levels of nutrients and metabolites for up to nine days of cultivation, representing an 11.1-fold increase and a maximum cell productivity of 422 cells/mL/h (1.0 × 10 5 viable cells/mL). ASCs maintained the immunophenotypic characteristics and mesodermal differentiation potential of both cell lines from different donors. The established protocol represents a more efficient and cost-effective method to obtain a high proliferation rate of ASCs in a microcarrier-based system, which is necessary for large-scale use in cell therapy, highlighting that the manipulation of critical parameters optimizes the ASCs production process.
(© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)
Databáze: MEDLINE
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