Stem cell engineering of striatal medium spiny neurons (MSNs) is a promising strategy to understand dis- eases affecting the striatum and for cell-replacement therapies in different neurological diseases. Protocols to generate cells from human pluripotent stem cells (PSCs) are scarce and how well they recapitulate the endogenous fetal cells remains poorly understood. We have developed a protocol that modulates cell seed- ing density and exposure to specific morphogens that generates authentic and functional D1- and D2-MSNs with a high degree of reproducibility in 25 days of differentiation. Single-cell RNA sequencing (scRNA-seq) shows that our cells can mimic the cell-fate acquisition steps observed in vivo in terms of cell type compo- sition, gene expression, and signaling pathways. Finally, by modulating the midkine pathway we show that we can increase the yield of MSNs. We expect that this protocol will help decode pathogenesis factors in striatal diseases and eventually facilitate cell-replacement therapies for Huntington’s disease (HD).

In vitro-derived medium spiny neurons recapitulate human striatal development and complexity at single-cell resolution / P. Conforti, V.D. Bocchi, I. Campus, L. Scaramuzza, M. Galimberti, T. Lischetti, F. Talpo, M. Pedrazzoli, A. Murgia, I. Ferrari, C. Cordiglieri, A. Fasciani, E. Arenas, D. Felsenfeld, G. Biella, D. Besusso, E. Cattaneo. - In: CELL REPORTS. METHODS. - ISSN 2667-2375. - 2:12(2022 Dec 19), pp. 100367.1-100367.18. [10.1016/j.crmeth.2022.100367]

In vitro-derived medium spiny neurons recapitulate human striatal development and complexity at single-cell resolution

P. Conforti
Primo
;
V.D. Bocchi;I. Campus;L. Scaramuzza;M. Galimberti;T. Lischetti;A. Murgia;I. Ferrari;G. Biella;D. Besusso
Penultimo
;
E. Cattaneo
Ultimo
2022

Abstract

Stem cell engineering of striatal medium spiny neurons (MSNs) is a promising strategy to understand dis- eases affecting the striatum and for cell-replacement therapies in different neurological diseases. Protocols to generate cells from human pluripotent stem cells (PSCs) are scarce and how well they recapitulate the endogenous fetal cells remains poorly understood. We have developed a protocol that modulates cell seed- ing density and exposure to specific morphogens that generates authentic and functional D1- and D2-MSNs with a high degree of reproducibility in 25 days of differentiation. Single-cell RNA sequencing (scRNA-seq) shows that our cells can mimic the cell-fate acquisition steps observed in vivo in terms of cell type compo- sition, gene expression, and signaling pathways. Finally, by modulating the midkine pathway we show that we can increase the yield of MSNs. We expect that this protocol will help decode pathogenesis factors in striatal diseases and eventually facilitate cell-replacement therapies for Huntington’s disease (HD).
Settore BIO/14 - Farmacologia
7PQCP-CSA13ECATT_M - European stem cell consortium for neural cell replacement, reprogramming and functional brain repair - CATTANEO, ELENA - 7PQCP-CSA - 7 Programma Quadro_Collaborative Project/Network/Coordination and Support Action - 2013
H20_RIA20ECATT_01 - Novel Strategies for Cell-based Neural Reconstruction (NSC-Reconstruct) - CATTANEO, ELENA - H20_RIA - Horizon 2020_Research & Innovation Action/Innovation Action - 2020
https://www.sciencedirect.com/science/article/pii/S2667237522002697
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/949402
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