Scale up manufacturing approach for production of human induced pluripotent stem cell-derived islets using Vertical Wheel® bioreactors
Nidheesh Dadheech, Mario Bermúdez de León, Zofia Czarnecka, Nerea Cuesta‐Gomez, Ila Tewari Jasra, Rena Pawlick, Braulio A. Marfil‐Garza, Sandhya Sapkota, Kevin Verhoeff, Haide Razavy, Perveen Anwar, Abhineet Singh, Nilanjan Ray, Doug O’ Gorman, Glen C. Jickling, James Lyon, Patrick E. MacDonald, A. M. James Shapiro
Abstract
Advanced protocols show potential for human stem cells (SC)-derived islets generation under planar (2D) alone or three-dimensional (3D) cultures, but show challenges in scalability, cell loss, and batch-to-batch consistency. This study explores Vertical Wheel (VW)® bioreactor suspension technology to differentiate islets from human induced pluripotent stem cells, achieving uniform, transcriptionally mature, and functional SC-islets. A 5x increase in scale from 0.1 L to 0.5 L reactors resulted in a 12-fold (15,005–183,002) increase in islet equivalent count (IEQ) without compromising islet structure. SC-islets show enriched β-cell composition (~63% CPPT + NKX6.1 + ISL1 + ), glucose responsive insulin release (3.9–6.1-fold increase), and reversed diabetes in STZ-treated mice. Single cell RNA sequencing and flowcytometry analysis confirmed transcriptional maturity and functional identity, similar to adult islets. Lastly, harvested SC-islet grafts demonstrate improved islet functionality and mature transcriptomic signatures. Overall, scale-up in VW® bioreactor technology enhances IEQ yield with minimal variability and reduced cell loss, offering a pathway for clinical-grade SC-islet production.