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Culturing of Retinal Pigment Epithelial Cells on an Ex Vivo Model of Aged Human Bruch's Membrane
Published on: April 12, 2018
Decreased uncoupling protein 2 expression in aging retinal pigment epithelial cells
1Department of Ophthalmology, the Second Affiliated Hospital of Xi'an Medical University; Ocular immunology and Inflammation Institute; Shaanxi Provincial Clinical Research Center for Ophthalmology, Xi'an 710038, Shaanxi Province, China.
Aim:
To analyze the expression of uncoupling protein 2 (UCP2) in retinal pigment epithelium (RPE) cells at the different human age, further explore the possible new target of RPE cells protection.
Methods:
Adult retinal pigment epithelial-19 (ARPE-19) cells and the primary RPE cells at the different age (9-20y, 50-55y, 60-70y, >70y) were cultured and harvested. The expression of UCP2 in these cells was detected by reverse transcription-polymerase chain reaction (RT-PCR), Western blot and confocal microscopy.
Results:
Cells from the donors more than 60y are larger and more fibroblastic in appearance compared to ARPE-19 cells and those primary cultures obtained from the younger individuals by using phase-contrast micrographs. Results of RT-PCR, Western blot and confocal microscopy all showed that UCP2 was highly expressed in ARPE-19 cells and in the younger primary cultured human RPE cells at the age of 9-20y and 50-55y, whereas lower expression of UCP2 was measured in the older primary cultured human RPE cells at the age more than 60y.
Conclusion:
Expression of UCP2 gene is decreased in aged RPE cells, promoting the lower ability of anti-oxidation in these cells. It is indicated that UCP2 gene might be a new target for protecting the cells from oxidative stress damage.
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