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Large-Scale Expansion of Human Liver Stem Cells Using Two Different Bioreactor Systems.

Jan Thorbow1, Andrea Strauch1, Viktoria Pfening1

  • 1Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, 35390 Giessen, Germany.

Bioengineering (Basel, Switzerland)
|July 27, 2024
PubMed
Summary

Scalable expansion of human liver stem cells (HLSCs) is crucial for cell therapy. This study optimized HLSC expansion in bioreactors, achieving sufficient viable cells for therapeutic use.

Keywords:
human liver stem cellskLalong-term cultivationmulti-plate bioreactoroxygen limitationscale-upstirred-tank bioreactor

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Area of Science:

  • Cell Biology
  • Biotechnology
  • Regenerative Medicine

Background:

  • Human liver stem cells (HLSCs) hold therapeutic potential.
  • Scalable and controlled expansion is essential for their clinical application.
  • Optimizing culture conditions and scale-up methods is critical.

Purpose of the Study:

  • To define optimal process parameters for HLSC expansion in 2D cultures.
  • To evaluate the scalability of HLSC expansion using multi-plate and stirred-tank bioreactors.
  • To ensure the viability, functionality, and undifferentiated state of scaled-up HLSCs.

Main Methods:

  • Conventional 2D culture optimization (seeding density, antibiotics, metabolites).
  • Scale-up in multi-plate bioreactor (Xpansion) and stirred-tank bioreactor (STR).
  • Process analytical technology for controlled bioreactor environments.

Main Results:

  • Optimal seeding density of 4000 cells cm-2 identified.
  • Lactate and ammonia concentrations were critical factors for HLSC expansion.
  • Bioreactor systems (XPN and STR) demonstrated successful scale-up, yielding sufficient viable HLSCs.
  • XPN bioreactor achieved higher cell density than standard T-flask cultures.

Conclusions:

  • HLSC expansion can be successfully scaled using bioreactor technology.
  • Optimized processes yield sufficient numbers of viable, functional, and undifferentiated HLSCs for therapeutic applications.
  • This work supports the clinical translation of HLSC-based therapies.