Process development and scale-up of pluripotent stem cell manufacturing
ShuoHao Huang, Azher Razvi, Zoe Anderson-Jenkins, Danylo Sirskyj, Ming Gong, Anne-Marie Lavoie, Gary Pigeau
Abstract
Human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs), together human pluripotent stem cells (PSCs), have tremendous potential for production of cellular therapies for regenerative medicine. The final therapeutic dose of differentiated PSCs varies according to application; however, most indications have production requirements that cannot be met by traditional static tissue culture methods. Stirred-tank reactor expansion of PSCs represents a scalable solution to meet this developing demand. Here we present the process development of a 10 L single-use stirred tank bioreactor platform with the ability to generate > 1010 PSCs per production run and its applicability across both hESC and hiPSC cell lines. Manufacturing advancements are also presented. High-density seed bank inoculation of the process is demonstrated to decouple the adherent tissue culture requirement and normalize the cellular process input, as well as shorten the time requirement of the suspension seed train. Process correlations of pH and dissolved oxygen to viable cell densities are presented to remove or minimize the sampling requirements during expansion. Automated downstream processing, with volume reduction and washing, is demonstrated with 80–90% cell recovery and >94% viability. Together, these works represent seminal steps in the development of a controlled, automated and defined manufacturing process for PSCs.