FULLY-AUTOMATED CULTIVATION OF IPSCS AND DIFFERENTIATION TO IMSCS USING THE AUTOCRAT SYSTEM

Abstract

Background & Aim

Although major developments in the field of regenerative medicine have been made, osteoarthritis (OA) remains one of the major illnesses of our day. Mesenchymal stem cells (MSCs) have been suggested as a potential therapy for OA. The clinical success of MSCs has been mixed with several successful phase I and II trials and some phase III trials questioning the efficacy of MSCs as a treatment for OA. These mixed results are partly is attributed to the high degree of biologic variability and lack of standardisation across the manufacturing process of MSCs. MSCs derived from induced pluripotent stem cells (iPSCs) might address some of the heterogeneity derived from donor-to-donor variability. On the manufacturing side, automation is a key technology to increase standardisation and improve iMSC manufacturing in cell quality and manufacturing efficiency.

Methods, Results & Conclusion

In the EU project AutoCRAT the systems Autostem and StemCellDiscovery are combined to a fully-automated system for the manufacturing of iPSCs, differentiation to iMSCs and chondrocytes, as well as MSC derived extracellular vesicles (EVs). The Autostem system was previously developed for automated bioreactor-based MSC expansion and subsequent fill-and-finish. For AutoCRAT it was adapted to allow manufacturing of iMSCs and iMSC derived extracellular vesicles (EVs). Similarly, the StemCellDiscovery was developed for plate-based MSC cultivation and microscopic evaluation. This system was adapted to allow for iPSC cultivation and differentiation of iPSCs to iMSCs and chondrocytes.

This talk reports on the evaluation of this automated solution using a direct comparison of automated versus manual cultivation of iMSCs and iPSCs in terms of efficiency and cell quality. More importantly, it compares manual and automated differentiation of iPSCs to iMSCs. This comparison clearly illustrates the automated system is highly capable in the cultivation of iPSCs and iMSCs. The morphology and fold expansion of iPSCs and iMSCs manufactured in an automated setting is comparable to the manual cultivation. The immunofluorescence and flow cytometry results confirm the automated system is capable of manufacturing high-quality iPSCs and iMSCs. Comparing automated and manual differentiation of iPSCs to iMSCs illustrates the capabilities of the AutoCRAT system in manufacturing of iPSC derived cell therapies. The data presented here highlights the importance of automation for the standardised and transparent manufacturing of cell therapies.