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Long‐term expansion of human induced pluripotent stem cells in a microcarrier‐based dynamic system
Author(s) -
Badenes Sara M,
Fernandes Tiago G,
Miranda Cláudia C,
PuschKlein Annette,
Haupt Simone,
Rodrigues Carlos AV,
Diogo Maria Margarida,
Brüstle Oliver,
Cabral Joaquim MS
Publication year - 2017
Publication title -
journal of chemical technology and biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.64
H-Index - 117
eISSN - 1097-4660
pISSN - 0268-2575
DOI - 10.1002/jctb.5074
Subject(s) - microcarrier , induced pluripotent stem cell , stem cell , cell culture , regenerative medicine , microbiology and biotechnology , cell , tissue engineering , cellular differentiation , chemistry , cell therapy , biomedical engineering , biology , biochemistry , embryonic stem cell , medicine , genetics , gene
BACKGROUND Human induced pluripotent stem ( hiPS ) cells provide a fascinating tool for exploring disease mechanisms, compound screening in pharmaceutical drug development, and might also represent a renewable source of cells for regenerative medicine applications. This requires increased cell quantities, generated under Good Manufacturing Practice‐compatible conditions in a scalable system. RESULTS A microcarrier‐based suspension culture was explored for scaling‐up hiPS cell expansion in serum‐free medium using synthetic peptide‐acrylate surface microcarriers, developed for long‐term support of hiPS cell self‐renewal. After a 7 day‐culture in a spinner flask, cells maintained their typical morphology, pluripotency‐associated marker expression and their differentiation capability. Envisaging improvement of the scalability of the culture, long‐term expansion on the microcarriers was attained using confluent microcarriers as the inoculum of successive spinner flask cultures. Importantly, bead‐to‐bead cell transfer allowed four consecutive sub‐culture procedures and a cumulative 241‐fold expansion was achieved within 15 days, leading to a total viable cell number of 3.3 × 10 8 cells. CONCLUSION This work is expected to enable the scale‐up of hiPS cell culture under defined conditions and potentially leading to the use of pluripotent stem cell derivatives in cell replacement therapies. © 2016 Society of Chemical Industry