Robotic high-throughput biomanufacturing and functional differentiation of human pluripotent stem cells
Carlos A. Tristan, Pinar Ormanoglu, Jaroslav Slamecka, Claire Malley, Pei‐Hsuan Chu, Vukasin M. Jovanovic, Yeliz Gedik, Yogita Jethmalani, Charles Bonney, Elena Barnaeva, John Braisted, Sunil K. Mallanna, Dorjbal Dorjsuren, Michael J. Iannotti, Ty C. Voss, Sam Michael, Anton Simeonov, Ilyas Singeç
Abstract
Efficient translation of human induced pluripotent stem cells (hiPSCs) requires scalable cell manufacturing strategies for optimal self-renewal and functional differentiation. Traditional manual cell culture is variable and labor intensive, posing challenges for high-throughput applications. Here, we established a robotic platform and automated all essential steps of hiPSC culture and differentiation under chemically defined conditions. This approach allowed rapid and standardized manufacturing of billions of hiPSCs that can be produced in parallel from up to 90 different patient- and disease-specific cell lines. Moreover, we established automated multi-lineage differentiation and generated functional neurons, cardiomyocytes, and hepatocytes. To validate our approach, we compared robotic and manual cell culture operations and performed comprehensive molecular and cellular characterizations (e.g., single-cell transcriptomics, mass cytometry, metabolism, electrophysiology) to benchmark industrial-scale cell culture operations toward building an integrated platform for efficient cell manufacturing for disease modeling, drug screening, and cell therapy.