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Production of definitive endoderm from human pluripotent stem cells

2021
Hannover Medical School, Hannover, Germany
Within this article, a detailed protocol for mass production of definitive endoderm (DE) cells from human pluripotent stem cells (hPSCs) in scalable and easy-to-handle suspension culture using a rotating Erlenmeyer flask or a stirred tank bioreactor is described. This protocol uses two different chemically defined and xeno-free media, and thereby is compatible with good manufacturing practice (GMP), and includes procedures for cryopreservation of obtained cells. The protocol allows for efficient DE cell production within 3 days and generates DE cells with >92% purity when using the bioreactor. The hPSC-derived DE cells can be differentiated into various endodermal lineages. This protocol should facilitate the flexible production of mature DE derivatives for physiologically relevant disease models, high-throughput drug screening, and toxicology testing, as well as cellular therapies.
Production and cryopreservation of definitive endoderm from human pluripotent stem cells under defined and scalable culture conditions
Ulrich Martin, Ruth Olmer
#1516
Added on: 08-08-2022
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