P06 | MODELING THE COMPLEXITY OF THE CENTRAL NERVOUS SYSTEM WITH PATIENT IPSC-DERIVED CEREBRAL ORGANOIDS

L. Carpinelli¹, D. Sibilia¹, P.N. Doronzio², F. Giacalone¹, C. Lisciandrello², E. Guadagni², A. Conte³, S. Lattante⁴, M. Magatti⁵, A. Silini⁵, Facility Biobanca⁶, Facility Citometria a Flusso⁶, L. Di Pietro^1|2, M. Sabatelli³, O. Parolini^1|2 | ¹Fondazione Policlinico Universitario A. Gemelli IRCSS, Rome, Italy; ²Dipartimento di Scienze della Vita e Sanità Pubblica, Università Cattolica del Sacro Cuore, Rome, Italy; ³Centro Clinico NEMO, Fondazione Policlinico Gemelli IRCCS, Rome, Italy; ⁴Department of Experimental Medicine, University of Salento, Lecce, Italy; ⁵Centro di Ricerca E. Menni, Fondazione Poliambulanza Istituto Ospedaliero, Brescia, Italy; ⁶Gemelli Science and Technology Park (GSTeP), Fondazione Policlinico Universitario A. Gemelli IRCSS, Rome, Italy

2D cultures poorly model tissue complexity. This study aims to develop a patient-iPSC-derived 3D model of the central nervous system - integrating neuronal and immune components - to study neurodegenerative diseases, specifically amyotrophic lateral sclerosis (ALS). ALS patients and healthy control fibroblasts were reprogrammed into iPSCs to generate liquid-cultured brain organoids (BOs), maintained for 50, 70, 80 and 100 days. To improve model complexity, BOs were stimulated with specific growth factors to induce oligodendrocytes differentiation. Microglial cells (generated from matched patient-derived iPSCs) were integrated. Morphometric analysis showed regular growth of both CTRL and ALS BOs over time, suggesting morphological stability. Immunofluorescence and gene expression analyses confirmed correct neuronal differentiation and spatial organization in both groups. Motor neurons and oligodendrocytes were detected. Microglia were effectively integrated into both CTRL and ALS BOs, displaying an amoeboid morphology in the latter group, suggesting a more activated state compared with CTRL BOs. This study establishes an optimized protocol for modeling a 3D platform suitable for investigating ALS.
Acknowledgements: Funded by the European Union – Next Generation EU – PNRR M6C2 – Investment 2.1 Enhancement and strengthening of biomedical research in the National Health Service – project PNRR-MCNT2-2023-12377338 – [PI: Ornella Parolini; Co-PI: Mario Sabatelli] – CUP:  C53C23001090007.