Long-term tripotent differentiation capacity of human neural stem (NS) cells in adherent culture.
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.
Sun
Yirui
Y
Wellcome Trust Centre for Stem Cell Research and Department of Biochemistry, University of Cambridge, Cambridge, UK.
Pollard
Steven
S
Conti
Luciano
L
Toselli
Mauro
M
Biella
Gerardo
G
Parkin
Georgina
G
Willatt
Lionel
L
Falk
Anna
A
Cattaneo
Elena
E
Smith
Austin
A
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
D015415:Biomarkers; D039842:Neural Cell Adhesion Molecule L1; D012794:Sialic Acids; C103469:polysialyl neural cell adhesion molecule; D049452:Green Fluorescent Proteins