Successful Use of Human AB Serum to Support the Expansion of Adipose Tissue-Derived Mesenchymal Stem/Stromal Cell in a Microcarrier-Based Platform

Author(s)

Moreira, Francisco ^[1] ; Mizukami, Amanda ^[2] ; de Souza, Lucas Eduardo Botelho ^[2] ; Cabral, Joaquim M. S. ^[2] ; da Silva, Claudia L. ^[2] ; Covas, Dimas T. ^[2] ; Swiech, Kamilla ^{[3, 2]}

Affiliation

^[1] Univ Lisbon, iBB Inst Bioengn & Biosci, Inst Super Tecn, Dept Bioengn, Lisbon - Portugal

^[2] Univ Sao Paulo, Ctr Cell Based Therapy, Reg Blood Ctr Ribeirao Preto, Sao Paulo - Brazil

^[3] Univ Sao Paulo, Sch Pharmaceut Sci Ribeirao Preto, Dept Pharmaceut Sci, Sao Paulo - Brazil

• Document type: Journal article
• Source: FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY; v. 8, APR 15 2020.
• Web of Science Citations: 1

Abstract

Mesenchymal stem/stromal cells (MSC) are promising candidates for cell-based therapies and for the promotion of tissue repair, hence the increase of clinical trials in a worldwide scale. In particular, adipose tissue-derived stem/stromal cells (AT MSC) present easy accessibility and a rather straightforward process of isolation, providing a clear advantage over other sources. The high demand of cell doses (millions of cells/kg), needed for infusion in clinical settings, requires a scalable and efficient manufacturing of AT MSC under xenogeneic(xeno)-free culture conditions. Here we describe the successful use of human AB serum {[}10%(v/v)] as a culture supplement, as well as coating substrate for the expansion of these cells in microcarriers using (i) a spinner flask and (ii) a 500-mL mini-bioreactor (Applikon(TM) Biotechnology). Cells were characterized by immunophenotype and multilineage differentiation potential. Upon an initial cell adhesion in the spinner flask of 35 +/- 2.5%, culture reached a maximal cell density of 2.6 +/- 0.1 x 10(5) at day 7, obtaining a 15 +/- 1-fold increase. The implementation of the culture in the 500-mL mini-bioreactor presented an initial cell adhesion of 22 +/- 5%, but it reached maximal cell density of 2.7 +/- 0.4 x 10(5) at day 7, obtaining a 27 +/- 8-fold increase. Importantly, in both stirred systems, cells retained their immunophenotype and multilineage differentiation potential (osteo-, chondro- and adipogenic lineages). Overall, the scalability of this microcarrier-based system presented herein is of major importance for the purpose of achieving clinically meaningful cell numbers. (AU)

• FAPESP's process: 13/08135-2 - CTC - Center for Cell-Based Therapy
• Grantee: Dimas Tadeu Covas
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 12/23228-4 - In vitro expansion of mesenchymal stromal cells and secretome characterization: therapeutic and biotechnological applications
• Grantee: Amanda Mizukami Martins
• Support Opportunities: Scholarships in Brazil - Doctorate