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  4. Automated real-time monitoring of human pluripotent stem cell aggregation in stirred tank reactors
 
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2019
Journal Article
Title

Automated real-time monitoring of human pluripotent stem cell aggregation in stirred tank reactors

Abstract
The culture of human induced pluripotent stem cells (hiPSCs) at large scale becomes feasible with the aid of scalable suspension setups in continuously stirred tank reactors (CSTRs). Innovative monitoring options and emerging automated process control strategies allow for the necessary highly defined culture conditions. Next to standard process characteristics such as oxygen consumption, pH, and metabolite turnover, a reproducible and steady formation of hiPSC aggregates is vital for process scalability. In this regard, we developed a hiPSC-specific suspension culture unit consisting of a fully monitored CSTR system integrated into a custom-designed and fully automated incubator. As a step towards cost-effective hiPSC suspension culture and to pave the way for flexibility at a large scale, we constructed and utilized tailored miniature CSTRs that are largely made from three-dimensional (3D) printed polylactic acid (PLA) filament, which is a low-cost material used in fused deposition modelling. Further, the monitoring tool for hiPSC suspension cultures utilizes in situ microscopic imaging to visualize hiPSC aggregation in real-time to a statistically significant degree while omitting the need for time-intensive sampling. Suitability of our culture unit, especially concerning the developed hiPSC-specific CSTR system, was proven by demonstrating pluripotency of CSTR-cultured hiPSCs at RNA (including PluriTest) and protein level.
Author(s)
Schwedhelm, Ivo
Zdzieblo, Daniela  
Appelt-Menzel, Antje  
Berger, Constantin  
Schmitz, Tobias
Schuldt, Bernhard
Franke, Andre
Müller, Franz-Josef
Pless, Ole
Schwarz, Thomas  
Wiedemann, Philipp
Walles, Heike
Fraunhofer-Institut für Silicatforschung ISC  
Hansmann, Jan  
Journal
Scientific Reports  
Open Access
DOI
10.1038/s41598-019-48814-w
Language
English
Fraunhofer-Institut für Silicatforschung ISC  
Keyword(s)
  • bioreactor

  • cell

  • suspension culture

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