Generation of primitive neural stem cells from human fibroblasts using a defined set of factors.
In mice, leukemia inhibitory factor (LIF)-dependent primitive neural stem cells (NSCs) have a higher neurogenic potential than bFGF-dependent definitive NSCs. Therefore, expandable primitive NSCs are required for research and for the development of therapeutic strategies for neurological diseases. There is a dearth of suitable techniques for the generation of human long-term expandable primitive NSCs. Here, we have described a method for the conversion of human fibroblasts to LIF-dependent primitive NSCs using a strategy based on techniques for the generation of induced pluripotent stem cells (iPSCs). These LIF-dependent induced NSCs (LD-iNSCs) can be expanded for >100 passages. Long-term cultured LD-iNSCs demonstrated multipotent neural differentiation potential and could generate motor neurons and dopaminergic neurons, as well as astrocytes and oligodendrocytes, indicating a high level of plasticity. Furthermore, LD-iNSCs easily reverted to human iPSCs, indicating that LD-iNSCs are in an intermediate iPSC state. This method may facilitate the generation of patient-specific human neurons for studies and treatment of neurodegenerative diseases.
Department of Reproductive Biology, National Center for Child Health and Development, Tokyo 157-8535, Japan miura-t@nihs.go.jp akutsu-h@ncchd.go.jp.
Takumi
T
Miura
Department of Reproductive Biology, National Center for Child Health and Development, Tokyo 157-8535, Japan.
Tohru
T
Sugawara
Department of Reproductive Biology, National Center for Child Health and Development, Tokyo 157-8535, Japan.
Atsushi
A
Fukuda
Department of Reproductive Biology, National Center for Child Health and Development, Tokyo 157-8535, Japan.
Ryo
R
Tamoto
Department of Reproductive Biology, National Center for Child Health and Development, Tokyo 157-8535, Japan.
Tomoyuki
T
Kawasaki
Department of Reproductive Biology, National Center for Child Health and Development, Tokyo 157-8535, Japan.
Akihiro
A
Umezawa
Department of Reproductive Biology, National Center for Child Health and Development, Tokyo 157-8535, Japan Department of Stem Cell Research, Fukushima Medical University, Fukushima 960-1295, Japan miura-t@nihs.go.jp akutsu-h@ncchd.go.jp.
Hidenori
H
Akutsu
Neural stem cell; Reprogramming; Stem cells