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

Author(s)

Simao, Vinicius Augusto ; Brand, Heloisa ; da Silveira-Antunes, Roseli Nunes ; Fukasawa, Josianne Thomazini ; Leme, Jaci ; Tonso, Aldo ; Ribeiro-Paes, Joao Tadeu

• Document type: Journal article
• Source: Biotechnology Letters; v. 45, n. 7, p. 24-pg., 2023-05-12.

Abstract

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-(R) 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 x 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 x 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. (AU)

• FAPESP's process: 14/03324-4 - Establishment of a bioprocess for the propagation of mesenchymal stem cells from adipose tissue
• Grantee: João Tadeu Ribeiro-Paes
• Support Opportunities: Regular Research Grants