TY - JOUR
T1 - Developmentally coordinated extrinsic signals drive human pluripotent stem cell differentiation toward authentic DARPP-32+ medium-sized spiny neurons
AU - Carri, Alessia Delli
AU - Onorati, Marco
AU - Lelos, Mariah J.
AU - Castiglioni, Valentina
AU - Faedo, Andrea
AU - Menon, Ramesh
AU - Camnasio, Stefano
AU - Vuono, Romina
AU - Spaiardi, Paolo
AU - Talpo, Francesca
AU - Toselli, Mauro
AU - Martino, Gianvito
AU - Barker, Roger A.
AU - Dunnett, Stephen B.
AU - Biella, Gerardo
AU - Cattaneo, Elena
PY - 2013
Y1 - 2013
N2 - 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.
AB - 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.
KW - DARPP-32 (PPP1R1B)
KW - Directed differentiation
KW - Human embryonic stem cells
KW - Huntington's disease
KW - Medium spiny neurons
KW - Striatal neuronal differentiation
UR - http://www.scopus.com/inward/record.url?scp=84871586691&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84871586691&partnerID=8YFLogxK
U2 - 10.1242/dev.084608
DO - 10.1242/dev.084608
M3 - Article
C2 - 23250204
AN - SCOPUS:84871586691
VL - 140
SP - 301
EP - 312
JO - Development (Cambridge)
JF - Development (Cambridge)
SN - 0950-1991
IS - 2
ER -