Stirred suspension bioreactors maintain na\xefve pluripotency of human pluripotent stem cells

Leili Rohani,Derek Toms,Breanna S Borys,Tiffany Dang,Ian A Lewis,Shiying Liu,Joyce W Li,Pooyan Naghsh,Ryan A Groves,Michael S Kallos,Tania So,Derrick E Rancourt,Heidrun Holland,Adiv A Johnson,Golsa Razian,Pranav Machiraju,Paul M K Gordon

doi:10.1038/s42003-020-01218-3

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.

Highlights

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
Considering that study[26], the robust expansion of mPSCs in bioreactors[8,9,10,11,16], and knowing that naïve Human pluripotent stem cells (hPSCs) resemble mPSCs, we report here that cell-state conversion significantly enhances hPSC bioprocessing in stirred suspension culture
Through transcriptomics, assessing the expression of epigenetic regulators, metabolomics, and cell-surface protein marker analyses, we show that the bioreactor environment helps maintain the naïve pluripotent state

Summary

Introduction

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. Through transcriptomic enrichment of mechanosensing 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. Pluripotency maintenance of this “high-suspension-yield” state was dependant on ERK inhibitory cytokine withdrawal from suspension culture This suggests that the cell’s interaction with its mechanical environment and media components plays a critical role in maintaining naïve-like pluripotency within the bioreactor. Considering that study[26], the robust expansion of mPSCs in bioreactors[8,9,10,11,16], and knowing that naïve hPSCs resemble mPSCs, we report here that cell-state conversion significantly enhances hPSC bioprocessing in stirred suspension culture.

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