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Study on cell culture and ability of proliferation and differentiation of rat retinal stem

Author: ZhangZuo
Tutor: ZhaoJiaLiang
School: Beijing Union Medical College
Course: Clinical
Keywords: retinal stem cell culture proliferation differentiation
CLC: R774.1
Type: PhD thesis
Year: 2009
Downloads: 11
Quote: 0
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Background:Ophthalmologists are still being unable to provide effective therapy for some diseases which damage retinal cells directly, such as glaucoma or retinal degeneration, etc. But the research of retinal stem cells (RSCs) brings hope to treat these diseases. Researchers have got some advance in the culture, proliferation and differentiation of RSCs, but there remains some problems. For example, there is no uniform culturing method, and the mechanism of the proliferation and differentiation of RSCs are still unclear. Further research in this realm will assist the future clinical application of RSCs.Objective:To culture rat RSCs using improved method, and to compare the proliferating and differentiating ability of RSCs from rats with different ages.Methods:Retinal neural layer mixed cells from rats of embryonic day15, postnatal within24hours and postnatal day5were cultured and passaged in culture medium which contained FBS. Immunofluorescence and RT-PCR were used to check RSCs and to semi-quantificate their ability of proliferating and differentiating, thus the comparison of this kind of ability of RSCs from rats with different ages could be carried out.Results:Rat RSCs were successfully cultured and passaged by the improved method. And as the mixed cells being passaged, RSCs proliferated and took advantage, and differentiated in the direction of retinal ganglion cells (RGCs). As being passaged, in cells from rats of embryonic day15, the Musashi and Nestin mRNA expression level held the line, while the Thy-1mRNA expression level grew up slowly; in cells from rats of postnatal within24hours, the Musashi mRNA expression level held the line, the Nestin mRNA expression level came down slowly, while the Thy-1mRNA expression level grew up; in cells from rats of postnatal day5, the Musashi and Nestin mRNA expression level came down, while the Thy-1mRNA expression level grew up.Conclusion:The improved method was stable and reliable in culturing and passaging RSCs, and could facilitate their differentiation in the direction of RGCs. RSCs from rats of embryonic day15had the strongest ability of proliferating and maintaining its primary property. And RSCs from rats of postnatal day5had the strongest ability of differentiating in the direction of retinal ganglion cells.

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CLC: > Medicine, health > Ophthalmology > Retina and optic nerve diseases > Retinal disease
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