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[Rat retinal ganglion cells in culture]
1Department of Ophthalmology, Second Affiliated Hospital, Hunan Medical University, Changsha 410011.
Objective:
To establish a culture system for retinal ganglion cells (RGCs) in order to lay a foundation for the experimental research in vitro.
Methods:
The retinae of 16 postnatal 2 - 3 day Sprague-Dawley rats were dissected into cell suspension with trypsin digestion. The cell suspension was implanted in 24 well culture plates with a cover slide 1 cm(2) in size and covered with murine tail collagen preplaced in each well (4 x 10(5) cells/well) and cultured under 37 degree C in an incubator with 5% CO(2). The cells were identified by immunocytochemical method with anti-rat Thy -1.1 monoclonal antibody after culture for 1, 3, 5 days, respectively, and the number of RGCs and its axon-growth percentage were counted in each 10-field high power (HP, 200 x) view under light microscope.
Results:
The RGCs cultured in murine tail collagen tissue grew very well. Some cells possessed axons and some axons connected in networks. The RGC number and its axon-growth percentage were (401 +/- 9) cells/10 HP and (25.34 +/- 0.72)% in 1-day-culture, (351 +/- 6) cells/10 HP and (35.16 +/- 2.22)% in 3-day-culture, (109 +/- 8) cells/10 HP and (69.84 +/- 0.97)% in 5-day-culture, respectively.
Conclusion:
RGCs can be cultured successfully and the murine tail collagen tissue is a good substratum for RGC survival in vitro.