Identifying and optimizing critical process parameters for large-scale manufacturing of iPSC derived insulin-producing β-cells

Haneen Yehya ^1,2, Alexandra Wells ¹, Michael Majcher ¹

⁰Trailhead Biosystems, 23215 Commerce Park, Beachwood, OH, 44122, USA.

Stem Cell Research & Therapy +

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November 10, 2024

View abstract on PubMed

Summary

Large-scale production of insulin-producing cells from induced pluripotent stem cells (iPSCs) is now possible using suspension bioreactors. This method enhances differentiation and offers potential for diabetes therapy.

Area Of Science

Biotechnology
Stem Cell Biology
Endocrinology

Background

Type 1 diabetes necessitates lifelong insulin therapy due to autoimmune destruction of pancreatic beta-cells.
Islet transplantation is a potential treatment but faces limitations in donor availability and immunosuppression requirements.
Induced pluripotent stem cells (iPSCs) offer a scalable source for generating functional beta-cells, but current differentiation methods are not optimized for large-scale suspension culture.

Purpose Of The Study

To develop and optimize a scalable suspension culture process for differentiating iPSCs into insulin-producing cells.
To investigate bioreactor scale-up parameters and their impact on iPSC differentiation for potential therapeutic applications.

Main Methods

Utilized an optimized High Dimensional Design of Experiments (HD-DoE) protocol for bioreactor scale-up.
Implemented a three-stage suspension manufacturing process transitioning from adherent to suspension culture in vertical wheel bioreactors.
Employed continuous bioreactor runs to analyze nutrient limitations and differentiation efficiency.

Main Results

A scalable three-stage suspension process was established using TB2 media for iPSC expansion.
Extended differentiation times and stage-wise optimization enhanced marker expression and maturation of iPSC-derived islet-like clusters.
Continuous bioreactors demonstrated metabolic shifts and a more beta-cell-like differentiation profile compared to control bioreactors. Cryopreserved aggregates maintained viability and insulin secretion capacity.

Conclusions

Increased stage duration and controlled media replenishment, leading to lactate accumulation, enhance the differentiation capacity of insulin-producing cells in large-scale suspension cultures.
The developed suspension culture process shows promise for the large-scale production of functional beta-cells for potential diabetes transplantation therapies.

Keywords:

Bioprocess development Bioreactor Diabetes DoE Human induced pluripotent stem cell Insulin producing cells Islets Optimization Pancreatic cells iPSCs β-cells