title: Human pluripotent stem cell differentiation into authentic striatal projection neurons.
abstract: 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 80 days 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.
authors.affiliation: Department of Biosciences and 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: Valentina
authors.initials: V
authors.lastname: Castiglioni
authors.forename: Andrea
authors.initials: A
authors.lastname: Faedo
authors.forename: Stefano
authors.initials: S
authors.lastname: Camnasio
authors.forename: Mauro
authors.initials: M
authors.lastname: Toselli
authors.forename: Gerardo
authors.initials: G
authors.lastname: Biella
authors.forename: Elena
authors.initials: E
authors.lastname: Cattaneo
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology; D061252:Feeder Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D051379:Mice; D017072:Neostriatum / Q000166:cytology*; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned