Sensitivity of human pluripotent stem cells to insulin precipitation induced by peristaltic pump-based medium circulation: Considerations on process development
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2018-10-09 13:10
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MASSAI, Diana, BOLESANI, Emiliano, DIAZ, Diana Robl, KROPP, Christina, KEMPF, Henning, HALLOIN, Caroline, MARTIN, Ulrich, BRANISTE, Tudor, ISU, Giuseppe, HARMS, Vanessa, MORBIDUCCI, Umberto, DRAGER, Gerald, ZWEIGERDT, Robert. Sensitivity of human pluripotent stem cells to insulin precipitation induced by peristaltic pump-based medium circulation: Considerations on process development. In: Scientific Reports, 2017, vol. 7, p. 0. ISSN 2045-2322. DOI: https://doi.org/10.1038/s41598-017-04158-x
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Scientific Reports
Volumul 7 / 2017 / ISSN 2045-2322

Sensitivity of human pluripotent stem cells to insulin precipitation induced by peristaltic pump-based medium circulation: Considerations on process development

DOI: https://doi.org/10.1038/s41598-017-04158-x

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Massai Diana1, Bolesani Emiliano1, Diaz Diana Robl1, Kropp Christina1, Kempf Henning1, Halloin Caroline1, Martin Ulrich1, Braniste Tudor12, Isu Giuseppe134, Harms Vanessa5, Morbiducci Umberto6, Drager Gerald15, Zweigerdt Robert1
 
1 Hannover Medical School,
2 Technical University of Moldova,
3 University of Basel,
4 University Hospital of Basel,
5 Leibniz University Hannover,
6 Polytechnic of Turin
 
Disponibil în IBN: 2 februarie 2018


Rezumat

Controlled large-scale production of human pluripotent stem cells (hPSCs) is indispensable for their envisioned clinical translation. Aiming at advanced process development in suspension culture, the sensitivity of hPSC media to continuous peristaltic pump-based circulation, a well-established technology extensively used in hydraulically-driven bioreactors, was investigated. Unexpectedly, conditioning of low protein media (i.e. E8 and TeSR-E8) in a peristaltic pump circuit induced severe viability loss of hPSCs cultured as aggregates in suspension. Optical, biochemical, and cytological analyses of the media revealed that the applied circulation mode resulted in the reduction of the growth hormone insulin by precipitation of micro-sized particles. Notably, in contrast to insulin depletion, individual withdrawal of other medium protein components (i.e. bFGF, TGFβ1 or transferrin) provoked minor reduction of hPSC viability, if any. Supplementation of the surfactant glycerol or the use of the insulin analogue Aspart did not overcome the issue of insulin precipitation. In contrast, the presence of bovine or human serum albumin (BSA or HSA, respectively) stabilized insulin rescuing its content, possibly by acting as molecular chaperone-like protein, ultimately supporting hPSC maintenance. This study highlights the potential and the requirement of media optimization for automated hPSC processing and has broad implications on media development and bioreactor-based technologies.