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

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.