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A xeno‐free microcarrier‐based stirred culture system for the scalable expansion of human mesenchymal stem/stromal cells isolated from bone marrow and adipose tissue
Author(s) -
Carmelo Joana G.,
FernandesPlatzgummer Ana,
Diogo Maria Margarida,
da Silva Cláudia Lobato,
Cabral Joaquim M. S.
Publication year - 2015
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201400586
Subject(s) - microcarrier , mesenchymal stem cell , stromal cell , stem cell , microbiology and biotechnology , bone marrow , bioreactor , adipose tissue , cell culture , cell therapy , biomedical engineering , chemistry , immunology , cell , biology , medicine , cancer research , biochemistry , genetics , organic chemistry
Human mesenchymal stem/stromal cells (MSC) are promising candidates for cell‐based therapies and the development of microcarrier‐based cultures in scalable bioreactors with well‐defined xenogeneic‐free components represent important milestones towards the clinical‐scale production of these cells. In this work, we optimized our previously developed xeno‐free microcarrier‐based system for the scalable expansion of human MSC isolated from bone marrow (BM MSC) and adipose‐derived stem/stromal cells (ASC). By adapting the agitation/feeding protocol at the initial cell seeding/cultivation stage in spinner flasks, we were able to maximize cell expansion rate and final cell yield. Maximal cell densities of 3.6 × 10 5 and 1.9 × 10 5 cells/mL were obtained for BM MSC (0.60 ± 0.04 day –1 ) and ASC (0.9 ± 0.1 day –1 ) cultures, upon seven and eight days of cultivation, respectively. Ready‐to‐use microcarriers Synthemax® II and Enhanced Attachment® supported identical expansion performance of BM MSC, turning those effective alternatives to the pre‐coated plastic microcarriers used in our xeno‐free scalable culture system. Importantly, expanded MSC maintained their immunophenotype and multilineage differentiation potential. Moreover, secretome analysis suggested a priming effect of stirred culture conditions on cytokine production by MSC. This culture system yielded considerable final cell densities that can be scaled‐up to controlled large‐scale bioreactors allowing a more efficient, safe and cost‐effective MSC production for clinical settings.