title: Stem cells expanded from the human embryonic hindbrain stably retain regional specification and high neurogenic potency.
abstract: Stem cell lines that faithfully maintain the regional identity and developmental potency of progenitors in the human brain would create new opportunities in developmental neurobiology and provide a resource for generating specialized human neurons. However, to date, neural progenitor cultures derived from the human brain have either been short-lived or exhibit restricted, predominantly glial, differentiation capacity. Pluripotent stem cells are an alternative source, but to ascertain definitively the identity and fidelity of cell types generated solely in vitro is problematic. Here, we show that hindbrain neuroepithelial stem (hbNES) cells can be derived and massively expanded from early human embryos (week 5-7, Carnegie stage 15-17). These cell lines are propagated in adherent culture in the presence of EGF and FGF2 and retain progenitor characteristics, including SOX1 expression, formation of rosette-like structures, and high neurogenic capacity. They generate GABAergic, glutamatergic and, at lower frequency, serotonergic neurons. Importantly, hbNES cells stably maintain hindbrain specification and generate upper rhombic lip derivatives on exposure to bone morphogenetic protein (BMP). When grafted into neonatal rat brain, they show potential for integration into cerebellar development and produce cerebellar granule-like cells, albeit at low frequency. hbNES cells offer a new system to study human cerebellar specification and development and to model diseases of the hindbrain. They also provide a benchmark for the production of similar long-term neuroepithelial-like stem cells (lt-NES) from pluripotent cell lines. To our knowledge, hbNES cells are the first demonstration of highly expandable neuroepithelial stem cells derived from the human embryo without genetic immortalization.
authors.affiliation: Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge CB21QR, United Kingdom.
authors.forename: Jignesh
authors.initials: J
authors.lastname: Tailor
authors.forename: Raja
authors.initials: R
authors.lastname: Kittappa
authors.forename: Ketty
authors.initials: K
authors.lastname: Leto
authors.forename: Monte
authors.initials: M
authors.lastname: Gates
authors.forename: Melodie
authors.initials: M
authors.lastname: Borel
authors.forename: Ole
authors.initials: O
authors.lastname: Paulsen
authors.forename: Sonia
authors.initials: S
authors.lastname: Spitzer
authors.forename: Ragnhildur Thora
authors.initials: RT
authors.lastname: Karadottir
authors.forename: Ferdinando
authors.initials: F
authors.lastname: Rossi
authors.forename: Anna
authors.initials: A
authors.lastname: Falk
authors.forename: Austin
authors.initials: A
authors.lastname: Smith
mesh_terms: D000818:Animals; D016380:Brain Tissue Transplantation / Q000379:methods; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D049109:Cell Proliferation / Q000187:drug effects*; D002531:Cerebellum / Q000166:cytology; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000166:cytology*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D061252:Feeder Cells; D005260:Female; D005333:Fetus / Q000166:cytology*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006801:Humans; D008297:Male; D051379:Mice; D058953:Neural Stem Cells / Q000166:cytology*; D046569:Neuroepithelial Cells / Q000166:cytology*; D061251:Primary Cell Culture; D051381:Rats; D017207:Rats, Sprague-Dawley; D012249:Rhombencephalon / Q000166:cytology*; D033581:Stem Cell Transplantation / Q000379:methods
chemical_list: D016222:Fibroblast Growth Factor 2; D004815:Epidermal Growth Factor