LARGE SCALE MANUFACTURING AND POTENCY ASSAY DEVELOPMENT FOR HMSCS IN REGENERATIVE MEDICINE

ABSTRACT

Background & Aim: A consortium of leading human mesenchymal stem/stromal cell (hMSC) therapy developers, Sentien Biotechnologies, GenCure Biomanufacturing and RoosterBio, are developing a large-scale hMSC biomanufacturing process with a deep Quality focus, culminating in a potency assay qualified with human clinical samples. Methods, Results & Conclusion: A Xeno-Free (XF), fed-batch, microcarrier-based bioreactor process for hMSC manufacturing had been developed and optimized [1], and scaled to a 50L process, with demonstrated comparability between the hMSC critical quality attributes (CQAs) from the bioreactor process and from 2D control cells of similar population doubling (PDL) [2]. Based on this previous process development, GenCure and RoosterBio are leading the first stage of biomanufacturing through upstream (2D seed train & bioreactor expansion) and downstream process (continuous counterflow centrigufation, formulation & fill, and cryopreservation). Bioreactor runs at the 50 L scale were performed using the most commonly used hMSC sources in regenerative medicine bone marrow (BM-MSCs), umbilical cord (UC-MSCs) and adipose (AD-MSCs). Critical process parameters (CPPs) are defined and critical quality attributes (CQAs) of the harvested cell product are characterized. An initial production run produced over 33billion BM-MSCs that passed the ISCT minimal criteria for MSCs. The subsequent expansion of UC- and AD-MSCs will be presented. Sentien's proprietary platform was used to ask questions on how the MSCs reacted to different stimuli with results showing that BM-MSCs were able to sense and respond with different secretomes to inflammatory stimuli. Our data also showed that BMMSCs induced changes in CD4, CD8 and CD19 cells and significantly reduced TNF-a levels in activated PBMCs, demonstrating their immunomodulatory capabilities. The same assays will be performed using the resulting MSCs from umbilical cord and adipose tissue expanded in the 50L bioreactor. Sentien, which has treated 16 subjects with acute kidney injury (AKI, open IND) and is currently running a trial in severe COVID-19 patients with AKI, will contribute biomarker data from the clinical-scale bioreactor and patient samples. Testing the in vitro developed potency hypothesis against the clinical samples will form the basis of a true potency assay. Through this work, the consortium will develop a generalized quality framework for large scale MSC manufacturing and potency assay development for broad use in regenerative medicine.