1997 Fiscal Year Final Research Report Summary
Immortalization of rat midbrain progenitor cells for dopamine neurons
| Project/Area Number |
08671578
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Cerebral neurosurgery
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
HOSHIMARU Minoru Kyoto University Graduate School of Medicine, Associate Professor, 医学研究科, 講師 (70211539)
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| Project Period (FY) |
1996 – 1997
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| Keywords | Parkinson's disease / dopamine neuron / retrovirus / sonic hedgehog / v-myc / tyrosine hydroxylase |
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| Research Abstract |
A regulatable retroviral vector LINXv-myc, in which the v-myc transgene is transcribed in a tetracycline-regulated fashion and neomycin phosphotransferase is expressed constitutively, was used for conditional immortalization of embryonic rat midbrain neural cells. A pooled population of cells which were selected for stable transfectants after infection with this retrovirus, stopped proliferating, extended processes, and induced GFAP after suppression of v-myc expression with tetracycline. These cells was subsequently infected with another retroviral vector which expresses 20 kDa amino-terminal cleavage product of Sonic hedgehog (SHH-N) and histidinol dehydrogenase and selected. A clone which was resistent to L-histidinol, A1, also extended processes and induced GFAP after the treatment of tetracycline and secreted SHH-N.Co-culture with the differentiated A1 cells induced TH-positive cells in dissociated cell culture derived from midbrain of embryonic day 11 (E11) rats. The differentiated A1 cells exhibited a beneficial effect also on differentiation of E12 and E13 ventral midbrain cells. The A1 cell line may be useful to enrich dopaminergic neurons in a cell suspension graft.
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