title: Long-term tripotent differentiation capacity of human neural stem (NS) cells in adherent culture.
abstract: Stem cell lines that provide a renewable and scaleable supply of central nervous system cell types would constitute an invaluable resource for basic and applied neurobiology. Here we describe the generation and long-term expansion of multiple human foetal neural stem (NS) cell lines in monolayer culture without genetic immortalization. Adherent human NS cells are propagated in the presence of epidermal growth factor (EGF) and fibroblast growth factor 2 (FGF2), under which conditions they stably express neural precursor markers and exhibit negligible differentiation into neurons or glia. However, they produce astrocytes, oligodendrocytes, and neurons upon exposure to appropriate differentiation factors. Single cell cloning demonstrates that human NS cells are tripotent. They retain a diploid karyotype and constant neurogenic capacity after over 100 generations. In contrast to human neurospheres, we observe no requirement for the cytokine leukaemia inhibitory factor (LIF) for continued expansion of adherent human NS cells. Human NS cells can be stably transfected to provide reporter lines and readily imaged in live monolayer cultures, creating the potential for high content genetic and chemical screens.
authors.affiliation: Wellcome Trust Centre for Stem Cell Research and Department of Biochemistry, University of Cambridge, Cambridge, UK.
authors.forename: Yirui
authors.initials: Y
authors.lastname: Sun
authors.forename: Steven
authors.initials: S
authors.lastname: Pollard
authors.forename: Luciano
authors.initials: L
authors.lastname: Conti
authors.forename: Mauro
authors.initials: M
authors.lastname: Toselli
authors.forename: Gerardo
authors.initials: G
authors.lastname: Biella
authors.forename: Georgina
authors.initials: G
authors.lastname: Parkin
authors.forename: Lionel
authors.initials: L
authors.lastname: Willatt
authors.forename: Anna
authors.initials: A
authors.lastname: Falk
authors.forename: Elena
authors.initials: E
authors.lastname: Cattaneo
authors.forename: Austin
authors.initials: A
authors.lastname: Smith
mesh_terms: D001253:Astrocytes / Q000166:cytology*; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D002999:Clone Cells; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005333:Fetus / Q000166:cytology; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D039842:Neural Cell Adhesion Molecule L1 / Q000378:metabolism; D009474:Neurons / Q000166:cytology*; D009836:Oligodendroglia / Q000166:cytology*; D012794:Sialic Acids / Q000378:metabolism; D014162:Transfection
chemical_list: D015415:Biomarkers; D039842:Neural Cell Adhesion Molecule L1; D012794:Sialic Acids; C103469:polysialyl neural cell adhesion molecule; D049452:Green Fluorescent Proteins