I038 Clinical grade production of large-scale neural progenitor cells for Huntington’s disease treatment

<h3>Background</h3> Cell therapy aims to revolutionize medical practice by offering cures for severe diseases. This is achieved through the utilization of human pluripotent stem cell (hPSC) lines, which undergo controlled differentiation before being transplanted into patients. A significant challenge lies in developing safe, robust, and controlled differentiation protocols, requiring the adaptation of research-only reagents to good manufacturing practice (GMP) standards. Cell-based therapies for Huntington’s disease focus on restoring neuronal connectivity and function by replacing the affected neurons, the striatal projection neurons (SPNs). <h3>Aim</h3> This work aims to develop a robust and reproducible GMP-compliant protocol for the maintenance and differentiation of hPSCs into neural progenitor cells (NPCs) suitable for clinical applications. <h3>Methods</h3> We build on a recently described differentiation protocol for the maintenance and differentiation of hPSCs into NPCs, replacing products that do not comply with GMP standards with GMP-graded ones. Furthermore, we apply stringent quality controls to ensure product safety and effectiveness. <h3>Results</h3> We demonstrate that hPSCs maintain their morphology and intrinsic pluripotent properties under GMP-compliant culture and scaling conditions. We determine cell adhesion rates to VTN-N to predict seeding cell numbers and observe decreased scaling efficiency with flasks larger than T75. During cryopreservation, controlled-rate freezing (CRF) improved recovery rates. Upon differentiation, NPCs exhibited markers of SPN progenitors and functionality upon maturation. <h3>Conclusion</h3> Overall, we confirm the feasibility of deriving and differentiating clinical-grade hPSC lines into NPCs under GMP-compliant conditions. <h3>Reference</h3> Comella-Bolla A, Orlandi JG, Miguez A, <i>et al.</i> Human Pluripotent Stem Cell-Derived Neurons Are Functionally Mature In Vitro and Integrate into the Mouse Striatum Following Transplantation. <i>Mol Neurobiol</i>. 2020;<b>57</b>(6):2766-2798. This study was supported by grants from the Ministerio de Ciencia e Innovación (PID2021-126961OB-I00; and PDC2022-133783-I00); Instituto de Salud Carlos III, Ministerio de Ciencia e Innovación and European Regional Development Fund (ERDF A way of making Europe) (Red de Terapias Avanzadas, RD21/0017/0020); European Union NextGenerationEU/PRTR; and Generalitat de Catalunya (2021 SGR 01094), Spain. The project was also supported by European Union grant from the European Innovation Council (EIC) grant agreement 101047099 – 4-DBR.