High density bioprocessing of human pluripotent stem cells by metabolic control and in silico modeling

Stem Cells Translational Medicine

Stefan Kalies¹  Michael Peitz²  Yannik Breitkreuz²  Oliver Brüstle²  Christina Kropp³  Caroline Halloin³  Annika Franke³  Clara‐Milena Farr³  Alexandra Haase³  Ulrich Martin³  Robert Zweigerdt³  Felix Manstein³  Ruth Olmer³  Wiebke Triebert³  Kevin Ullmann³  Anais Sahabian³ 

[1] Institute of Quantum Optics, Leibniz University Hannover Hannover Germany;Institute of Reconstructive Neurobiology, University of Bonn Medical Faculty & University Hospital Bonn Bonn Germany;Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO) Germany;

关键词: high density culture;    human pluripotent stem cells;    in silico process modeling;    process scale‐up;    stirred tank bioreactor;    suspension culture;   

DOI  :  10.1002/sctm.20-0453

来源: DOAJ

【 摘 要 】

Abstract To harness the full potential of human pluripotent stem cells (hPSCs) we combined instrumented stirred tank bioreactor (STBR) technology with the power of in silico process modeling to overcome substantial, hPSC‐specific hurdles toward their mass production. Perfused suspension culture (3D) of matrix‐free hPSC aggregates in STBRs was applied to identify and control process‐limiting parameters including pH, dissolved oxygen, glucose and lactate levels, and the obviation of osmolality peaks provoked by high density culture. Media supplements promoted single cell‐based process inoculation and hydrodynamic aggregate size control. Wet lab‐derived process characteristics enabled predictive in silico modeling as a new rational for hPSC cultivation. Consequently, hPSC line‐independent maintenance of exponential cell proliferation was achieved. The strategy yielded 70‐fold cell expansion in 7 days achieving an unmatched density of 35 × 106 cells/mL equivalent to 5.25 billion hPSC in 150 mL scale while pluripotency, differentiation potential, and karyotype stability was maintained. In parallel, media requirements were reduced by 75% demonstrating the outstanding increase in efficiency. Minimal input to our in silico model accurately predicts all main process parameters; combined with calculation‐controlled hPSC aggregation kinetics, linear process upscaling is also enabled and demonstrated for up to 500 mL scale in an independent bioreactor system. Thus, by merging applied stem cell research with recent knowhow from industrial cell fermentation, a new level of hPSC bioprocessing is revealed fueling their automated production for industrial and therapeutic applications.

【 授权许可】

Unknown