Continuous WNT Control Enables Advanced hPSC Cardiac Processing and Prognostic Surface Marker Identification in Chemically Defined Suspension Culture

Publication date: Available online 25 July 2019Source: Stem Cell ReportsAuthor(s): Caroline Halloin, Kristin Schw.e, Wiebke Löbel, Annika Franke, Monika Szepes, Santoshi Biswanath, Stephanie Wunderlich, Sylvia Merkert, Natalie Weber, Felix Osten, Jeanne de la Roche, Felix Polten, Kai Wollert, Theresia Kraft, Martin Fischer, Ulrich Martin, Ina Gruh, Henning Kempf, Robert ZweigerdtSummaryAiming at clinical translation, robust directed differentiation of human pluripotent stem cells (hPSCs), preferentially in chemically defined conditions, is a key requirement. Here, feasibility of suspension culture based hPSC-cardiomyocyte (hPSC-CM) production in low-cost, xeno-free media compatible with good manufacturing practice standards is shown. Applying stirred tank bioreactor systems at increasing dimensions, our advanced protocol enables routine production of about 1 million hPSC-CMs/mL, yielding ∼1.3 × 108 CM in 150 mL and ∼4.0 × 108 CMs in 350–500 mL process scale at>90% lineage purity. Process robustness and efficiency is ensured by uninterrupted chemical WNT pathway control at early stages of differentiation and results in the formation of almost exclusively ventricular-like CMs. Modulated WNT pathway regulation also revealed the previously unappreciated role of ROR1/CD13 as superior surrogate markers for predicting cardiac differentiation efficiency as soon as 72 h of differentiation. This monitoring strategy facilitates process upscaling and controlled mass produc...
Source: Stem Cell Reports - Category: Stem Cells Source Type: research