Journal
CELLS
Volume 8, Issue 12, Pages -Publisher
MDPI
DOI: 10.3390/cells8121571
Keywords
pluripotent stem cells; definitive endoderm differentiation; suspension culture; multi-lineage potential; cryopreservation
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Funding
- German Research Foundation, Cluster of Excellence REBIRTH [DFG EXC62/3]
- German Center for Lung Research [BREATH 82DZL002A1]
- R2N, Federal State of Lower Saxony [74ZN1574]
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For the production and bio-banking of differentiated derivatives from human pluripotent stem cells (hPSCs) in large quantities for drug screening and cellular therapies, well-defined and robust procedures for differentiation and cryopreservation are required. Definitive endoderm (DE) gives rise to respiratory and digestive epithelium, as well as thyroid, thymus, liver, and pancreas. Here, we present a scalable, universal process for the generation of DE from human-induced pluripotent stem cells (hiPSCs) and embryonic stem cells (hESCs). Optimal control during the differentiation process was attained in chemically-defined and xeno-free suspension culture, and high flexibility of the workflow was achieved by the introduction of an efficient cryopreservation step at the end of DE differentiation. DE aggregates were capable of differentiating into hepatic-like, pancreatic, intestinal, and lung progenitor cells. Scale-up of the differentiation process using stirred-tank bioreactors enabled production of large quantities of DE aggregates. This process provides a useful advance for versatile applications of DE lineages, in particular for cell therapies and drug screening.
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