Establishment of an epidermal growth factor-dependent, multipotent neural precursor cell line.
We have established a multipotent clonal cell line, named MEB5, from embryonic mouse forebrains after the infection of a retrovirus carrying E7 oncogene of human papillomavirus type 16. MEB5 cells proliferated in serum-free, epidermal growth factor (EGF)-supplemented medium. They expressed markers for neural precursor cells (nestin, A2B5, and RC1) and did not express markers for neurons (class III beta-tubulin), astrocytes (glial fibrillary acidic protein), and oligodendrocytes (galactocerebroside). MEB5 cells were stably maintained in an undifferentiated state with a diploid karyotype in the presence of EGF. When they were deprived of EGF, about 50% of the cells died due to apoptosis within 24 h. The remaining cells differentiated into neurons, astrocytes, or oligodendrocytes within 2 wk. The newly developed cells with neuronal morphology were immunoreactive for gamma-aminobutyric acid and exhibited neuronal electrophysiological properties. When MEB5 cells were treated with leukemia inhibitory factor for 7 d, they were induced to differentiate exclusively into astrocytes. These results indicate that MEB5 is a cell line with characteristics of EGF-dependent, multipotent neural precursor cells. This cell line should provide a good model system to study the mechanisms of survival, proliferation, and differentiation of the multipotent precursor cells in the central nervous system.
Institute for Fermentation, Osaka, Japan.
Y
Y
Nakagaito
M
M
Satoh
H
H
Kuno
T
T
Iwama
M
M
Takeuchi
A
A
Hakura
T
T
Yoshida
D000818:Animals; D001253:Astrocytes / Q000166:cytology; D002454:Cell Differentiation / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002461:Cell Line, Transformed; D002470:Cell Survival / Q000502:physiology; D016895:Culture Media, Serum-Free; D004815:Epidermal Growth Factor / Q000502:physiology*; D051379:Mice; D008809:Mice, Inbred C3H; D009856:Oncogene Proteins, Viral / Q000235:genetics; D027383:Papillomaviridae / Q000235:genetics; D050725:Papillomavirus E7 Proteins; D010641:Phenotype; D016548:Prosencephalon / Q000166:cytology* / Q000196:embryology / Q000821:virology; D012190:Retroviridae / Q000235:genetics
D016895:Culture Media, Serum-Free; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; C059731:oncogene protein E7, Human papillomavirus type 16; D004815:Epidermal Growth Factor