Scale-down optimization of a robust, parallelizable human induced pluripotent stem cell bioprocess for...

Area of Science:

• Biotechnology
• Regenerative Medicine
• Stem Cell Biology

Background:

• Clinically relevant quantities of high-quality human induced pluripotent stem cells (hiPSCs) are crucial for regenerative medicine therapies.
• Limited understanding of phenotype-governing networks and high costs hinder clinical trial efficacy and study throughput.

Purpose of the Study:

• To develop an optimized, high-throughput strategy for expanding hiPSCs at a research scale (<20 mL).
• To enable statistically rigorous studies for advancing hiPSC-based clinical manufacturing.

Main Methods:

• Assessed single-cell inoculation and aggregate preformation for hiPSC proliferation.
• Modeled aggregate characteristics in relation to agitation rates.
• Quantified pluripotency markers and performed teratoma assays.

Main Results:

• Achieved tunable control over hiPSC expansion with fold expansion comparable to commercial systems.
• Demonstrated functional pluripotency confirmed by marker quantification and teratoma assays.
• Established a scalable protocol for accelerating hiPSC research.

Conclusions:

• The optimized strategy enables high-throughput hiPSC expansion, reducing costs and increasing study throughput.
• This work is a significant step towards elucidating links between hiPSC phenotype and derivative functionality.
• Facilitates statistically rigorous studies for advancing hiPSC-based therapies in clinical manufacturing.