|Budget Amount *help
¥2,200,000 (Direct Cost : ¥2,200,000)
Fiscal Year 1997 : ¥600,000 (Direct Cost : ¥600,000)
Fiscal Year 1996 : ¥1,600,000 (Direct Cost : ¥1,600,000)
Retinal neurons have been maitained in long-term dissociated cell cultures. Cell type-specific markers such rhodopsin, S-antigen, neurofilament and MAP 2 were positively stained. Neurites of ganglion cells, which grew on Mueller cells, elongated up to 500 mum. These results suggest that retinal-Mueller cell interaction might play one of essential roles in survival and regeneration.
In this project, we isolated total RNA from cultured retinal cells, especially Mueller cells. Neurotrophic factors, such as nerve growth factor (NGF) , brain-derived neurotrophic factor (BDNF) , glial cell line-derived neurotrophic factor (GDNF) , were evaluated by reverse transcription polymerase chain reaction (RT-PCR). Messenger RNA of NGF,BDNF,GDNF and IL-6 were expressed in cultured retinal cells. These demonstrsted that retinal cells might produce various neurotrophic factors for survival and regeneration of retinal neurons. In vitro and in vivo, condition medium were not enough to maintaine and grow retinal neurons. We found that more Mueller cells were necessary to get a large quantity of neurotrophic factors. And then, transformed Mueller cell lines were established using SV 40 large T antigen gene. Messenger RNA of NGF,BDNF,GDNF were also expressed in these cells. However, condition medium of Mueller cell line was still uneffective to maintain and grow retinal neurons. Therefore, we must continue to investigate this research.