title: Developmentally coordinated extrinsic signals drive human pluripotent stem cell differentiation toward authentic DARPP-32+ medium-sized spiny neurons.
abstract: Medium-sized spiny neurons (MSNs) are the only neostriatum projection neurons, and their degeneration underlies some of the clinical features of Huntington's disease. Using knowledge of human developmental biology and exposure to key neurodevelopmental molecules, human pluripotent stem (hPS) cells were induced to differentiate into MSNs. In a feeder-free adherent culture, ventral telencephalic specification is induced by BMP/TGFβ inhibition and subsequent SHH/DKK1 treatment. The emerging FOXG1(+)/GSX2(+) telencephalic progenitors are then terminally differentiated, resulting in the systematic line-independent generation of FOXP1(+)/FOXP2(+)/CTIP2(+)/calbindin(+)/DARPP-32(+) MSNs. Similar to mature MSNs, these neurons carry dopamine and A2a receptors, elicit a typical firing pattern and show inhibitory postsynaptic currents, as well as dopamine neuromodulation and synaptic integration ability in vivo. When transplanted into the striatum of quinolinic acid-lesioned rats, hPS-derived neurons survive and differentiate into DARPP-32(+) neurons, leading to a restoration of apomorphine-induced rotation behavior. In summary, hPS cells can be efficiently driven to acquire a functional striatal fate using an ontogeny-recapitulating stepwise method that represents a platform for in vitro human developmental neurobiology studies and drug screening approaches.
authors.affiliation: Center for Stem Cell Research, Università degli Studi di Milano, 20133 Milan, Italy.
authors.forename: Alessia
authors.initials: A
authors.lastname: Delli Carri
authors.forename: Marco
authors.initials: M
authors.lastname: Onorati
authors.forename: Mariah J
authors.initials: MJ
authors.lastname: Lelos
authors.forename: Valentina
authors.initials: V
authors.lastname: Castiglioni
authors.forename: Andrea
authors.initials: A
authors.lastname: Faedo
authors.forename: Ramesh
authors.initials: R
authors.lastname: Menon
authors.forename: Stefano
authors.initials: S
authors.lastname: Camnasio
authors.forename: Romina
authors.initials: R
authors.lastname: Vuono
authors.forename: Paolo
authors.initials: P
authors.lastname: Spaiardi
authors.forename: Francesca
authors.initials: F
authors.lastname: Talpo
authors.forename: Mauro
authors.initials: M
authors.lastname: Toselli
authors.forename: Gianvito
authors.initials: G
authors.lastname: Martino
authors.forename: Roger A
authors.initials: RA
authors.lastname: Barker
authors.forename: Stephen B
authors.initials: SB
authors.lastname: Dunnett
authors.forename: Gerardo
authors.initials: G
authors.lastname: Biella
authors.forename: Elena
authors.initials: E
authors.lastname: Cattaneo
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D002454:Cell Differentiation; D019070:Cell Lineage; D002470:Cell Survival; D017690:Cell Transplantation; D051937:Dopamine and cAMP-Regulated Phosphoprotein 32 / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000166:cytology; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005434:Flow Cytometry; D059330:GABAergic Neurons / Q000378:metabolism; D006801:Humans; D006816:Huntington Disease / Q000378:metabolism; D051379:Mice; D009474:Neurons / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D018408:Patch-Clamp Techniques; D039904:Pluripotent Stem Cells / Q000378:metabolism*; D017378:Quinolinic Acid / Q000494:pharmacology; D012313:RNA / Q000378:metabolism; D051381:Rats; D013234:Stem Cells / Q000166:cytology; D013997:Time Factors
chemical_list: D051937:Dopamine and cAMP-Regulated Phosphoprotein 32; D012313:RNA; D017378:Quinolinic Acid