Intracellular Measurement-Informed Multiscale Modeling for Scalable iPSC Manufacturing

Area of Science:

• Biotechnology and Bioprocessing
• Stem Cell Biology
• Metabolic Engineering

Background:

• Scalable manufacturing of human induced pluripotent stem cells (iPSCs) is critical for cell therapies.
• Existing 3D aggregate cultures show spatial and metabolic heterogeneity, hindering mechanistic understanding and predictive modeling.
• Laboratory-scale monolayer systems are more homogeneous but do not fully represent manufacturing conditions.

Purpose of the Study:

• To develop a multiscale mechanistic model for human induced pluripotent stem cells (iPSCs) that accounts for spatial and metabolic heterogeneity in 3D aggregate cultures.
• To link molecular, cellular, and macroscopic processes for improved mechanistic understanding and predictive metabolic modeling.
• To provide a quantitative foundation for scalable iPSC biomanufacturing.

Main Methods:

• Developed a modular multiscale mechanistic foundation model integrating extracellular dynamics, intracellular metabolic fluxes, and cellular redox states.
• Extended a monolayer kinetic network and coupled it with a biological systems-of-systems (Bio-SoS) multiscale model for aggregate cultures.
• Utilized systematic experiments including isotopic tracers, metabolite profiling, and two-photon optical redox imaging for model validation.

Main Results:

• The integrated framework successfully unified heterogeneous datasets across different culture configurations (monolayer and aggregate).
• The model enabled mechanistic interpretation of metabolic and redox responses in heterogeneous iPSC cultures.
• Demonstrated the capability to predict and understand metabolic behavior across various culture scales.

Conclusions:

• The developed multiscale model provides a quantitative foundation for understanding and optimizing scalable iPSC biomanufacturing.
• Accounting for spatial and metabolic heterogeneity is crucial for accurate modeling of iPSC aggregate cultures.
• This approach facilitates the advancement of cell therapies and regenerative medicines through improved manufacturing processes.