Establishment of culturing retinal ganglion cells and myelination

Research Project

Project/Area Number	01570983
Research Category	Grant-in-Aid for General Scientific Research (C)
Allocation Type	Single-year Grants
Research Field	Ophthalmology
Research Institution	University of Occupational and Environmental Health, Japan
Principal Investigator	TAKAHASHI Hiroshi UOEH, Dept Ophthalmol., Assistant Professor, 医学部, 講師 (00129511)
Co-Investigator(Kenkyū-buntansha)	MIYATA Hiroshi Keio Univ., Dept Ophthalmol., Instructor, 医学部, 助手 (60174190) FURUKAWA Hiroshi UOEH, Dept Anatomy, Assistant Professor, 医学部, 講師 (80131936)
Project Period (FY)	1989 – 1991
Project Status	Completed (Fiscal Year 1991)
Budget Amount *help	¥2,100,000 (Direct Cost: ¥2,100,000) Fiscal Year 1991: ¥300,000 (Direct Cost: ¥300,000) Fiscal Year 1990: ¥300,000 (Direct Cost: ¥300,000) Fiscal Year 1989: ¥1,500,000 (Direct Cost: ¥1,500,000)
Keywords	Retina / Ganglion cell / Oligodendrocyte / Myelin / Tissue culture
Research Abstract	Establishment of pure method of culturing retinal ganglion cells is very important for basic science and clinic because of revealing pathogenesis of demyelination, multiple sclerosis and developing transplantation of retinal neurons, especially ganglion cells. Enzyme digestion-Percoll density gradient, panning and filter paper methods were used for culturing pure retinal ganglion cells. We revealed these methods had some merits and some demerits. In the density grandient method, cells were separated three fractions, I to III. Large, middle and small cells were cultured, respectively. Large cells might be ganglion cells, and then they were surrounded with flat cells or Muller cells. Therefore, ganglion cells in fraction I were mixed with other cells. The panning method was very expensive and cells were characteristically pure, but the cell number was very low. After using filter paper method, purity of gangion cells was excellent, but the numbers were very low. Myelination was studied in co-culturing retinal explant and oligodendrocytes. These cells were twined around retinal neurites, and were attacted with neurites. These results suggest that neurites might be interacted with oligodendrocytes. However, silver staining was negative and myelin protein was not found.