naa a22 4500 510739377 CHVBK 20180411083022.0 cr unu---uuuuu 180411e20130801xx s 000 0 eng 10.1007/s12015-013-9441-8 doi (NATIONALLICENCE)springer-10.1007/s12015-013-9441-8 Human Pluripotent Stem Cell Differentiation into Authentic Striatal Projection Neurons [Elektronische Daten] [Alessia Delli Carri, Marco Onorati, Valentina Castiglioni, Andrea Faedo, Stefano Camnasio, Mauro Toselli, Gerardo Biella, Elena Cattaneo] Here we present the principles and steps of a protocol that we have recently developed for the differentiation of hES/iPS cells into the authentic human striatal projection medium spiny neurons (MSNs) that die in Huntington's Disease (HD). Authenticity is judged by the convergence of multiple features within individual cells. Our procedure lasts 80days and couples neural induction via BMP/TGF-β inhibition with exposure to the developmental factors sonic hedgehog (SHH) and dickkopf1 (DKK-1) to drive ventral telencephalic specification, followed by terminal differentiation [1]. Authenticity of the resulting neuronal population is monitored by the appearance of FOXG1+/GSX2+ progenitor cells of the lateral ganglionic eminence (LGE) at day 15-25 of differentiation, followed by appearance of CTIP2-, FOXP1- and FOXP2-positive cells at day 45. These precursor cells then mature into MAP2+/GABA+ neurons with 20% of them ultimately co-expressing the DARPP-32 and CTIP2 diagnostic markers and carrying electrophysiological properties expected for fully functional MSNs. The protocol is characterized by its replicability in at least three human pluripotent cell lines. Altogether this protocol defines a useful platform for in vitro developmental neurobiology studies, drug screening, and regenerative medicine approaches. Springer Science+Business Media New York, 2013 Striatal neuronal differentiation nationallicence Medium spiny neurons nationallicence DARPP-32 nationallicence Huntington's disease nationallicence Directed differentiation nationallicence Human embryonic stem cells nationallicence Human pluripotent stem cells nationallicence Delli Carri Alessia Department of Biosciences and Center for Stem Cell Research, Università degli Studi di Milano, 20133, Milan, Italy aut Onorati Marco Department of Biosciences and Center for Stem Cell Research, Università degli Studi di Milano, 20133, Milan, Italy aut Castiglioni Valentina Department of Biosciences and Center for Stem Cell Research, Università degli Studi di Milano, 20133, Milan, Italy aut Faedo Andrea Department of Biosciences and Center for Stem Cell Research, Università degli Studi di Milano, 20133, Milan, Italy aut Camnasio Stefano Department of Biosciences and Center for Stem Cell Research, Università degli Studi di Milano, 20133, Milan, Italy aut Toselli Mauro Department of Biology and Biotechnology, University of Pavia, 27100, Pavia, Italy aut Biella Gerardo Department of Biology and Biotechnology, University of Pavia, 27100, Pavia, Italy aut Cattaneo Elena Department of Biosciences and Center for Stem Cell Research, Università degli Studi di Milano, 20133, Milan, Italy aut Stem Cell Reviews and Reports Springer US; http://www.springer-ny.com 9/4(2013-08-01), 461-474 1550-8943 9:4<461 2013 9 12015 https://doi.org/10.1007/s12015-013-9441-8 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s12015-013-9441-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Delli Carri Alessia Department of Biosciences and Center for Stem Cell Research, Università degli Studi di Milano, 20133, Milan, Italy aut NATIONALLICENCE 700 1- Onorati Marco Department of Biosciences and Center for Stem Cell Research, Università degli Studi di Milano, 20133, Milan, Italy aut NATIONALLICENCE 700 1- Castiglioni Valentina Department of Biosciences and Center for Stem Cell Research, Università degli Studi di Milano, 20133, Milan, Italy aut NATIONALLICENCE 700 1- Faedo Andrea Department of Biosciences and Center for Stem Cell Research, Università degli Studi di Milano, 20133, Milan, Italy aut NATIONALLICENCE 700 1- Camnasio Stefano Department of Biosciences and Center for Stem Cell Research, Università degli Studi di Milano, 20133, Milan, Italy aut NATIONALLICENCE 700 1- Toselli Mauro Department of Biology and Biotechnology, University of Pavia, 27100, Pavia, Italy aut NATIONALLICENCE 700 1- Biella Gerardo Department of Biology and Biotechnology, University of Pavia, 27100, Pavia, Italy aut NATIONALLICENCE 700 1- Cattaneo Elena Department of Biosciences and Center for Stem Cell Research, Università degli Studi di Milano, 20133, Milan, Italy aut NATIONALLICENCE 773 0- Stem Cell Reviews and Reports Springer US; http://www.springer-ny.com 9/4(2013-08-01), 461-474 1550-8943 9:4<461 2013 9 12015 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer