Controlling expansion and cardiomyogenic differentiation of human pluripotent stem cells in scalable suspension culture

Henning Kempf, Ruth Olmer, Christina Kropp, Michael Rückert, Monica Jara-Avaca, Diana Robles-Diaz, Annika Franke, David A. Elliott, Daniel Wojciechowski, Martin Fischer, Angelica Roa Lara, George Kensah, Ina Gruh, Axel Haverich, Ulrich Martin, Robert Zweigerdt

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138 Citations (Scopus)

Abstract

To harness the potential of human pluripotent stem cells (hPSCs), an abundant supply of their progenies is required. Here, hPSC expansion as matrix-independent aggregates in suspension culture was combined with cardiomyogenic differentiation using chemical Wnt pathway modulators. A multiwell screen was scaled up to stirred Erlenmeyer flasks and subsequently to tank bioreactors, applying controlled feeding strategies (batch and cyclic perfusion). Cardiomyogenesis was sensitive to the GSK3 inhibitor CHIR99021 concentration, whereas the aggregate size was no prevailing factor across culture platforms. However, in bioreactors, the pattern of aggregate formation in the expansion phase dominated subsequent differentiation. Global profiling revealed a culture-dependent expression of BMP agonists/antagonists, suggesting their decisive role in cell-fate determination. Furthermore, metallothionein was discovered as a potentially stress-related marker in hPSCs. In 100 ml bioreactors, the production of 40 million predominantly ventricular-like cardiomyocytes (up to 85% purity) was enabled that were directly applicable to bioartificial cardiac tissue formation.

Original languageEnglish
Pages (from-to)1132-1146
Number of pages15
JournalStem Cell Reports
Volume3
Issue number6
DOIs
Publication statusPublished - 9 Dec 2014
Externally publishedYes

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