Stirred suspension bioreactors maintain naïve pluripotency of human pluripotent stem cells
Leili Rohani, Breanna S. Borys, Golsa Razian, Pooyan Naghsh, Shiying Liu, Adiv A. Johnson, Pranav Machiraju, Heidrun Holland, Ian A. Lewis, Ryan A. Groves, Derek Toms, Paul M. K. Gordon, Joyce W. Li, Tania So, Tiffany Dang, Michael S. Kallos, Derrick E. Rancourt
Abstract
Due to their ability to standardize key physiological parameters, stirred suspension bioreactors can potentially scale the production of quality-controlled pluripotent stem cells (PSCs) for cell therapy application. Because of differences in bioreactor expansion efficiency between mouse (m) and human (h) PSCs, we investigated if conversion of hPSCs, from the conventional "primed" pluripotent state towards the "naïve" state prevalent in mPSCs, could be used to enhance hPSC production. Through transcriptomic enrichment of mechano-sensing signaling, the expression of epigenetic regulators, metabolomics, and cell-surface protein marker analyses, we show that the stirred suspension bioreactor environment helps maintain a naïve-like pluripotent state. Our research corroborates that converting hPSCs towards a naïve state enhances hPSC manufacturing and indicates a potentially important role for the stirred suspension bioreactor's mechanical environment in maintaining naïve-like pluripotency.