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Y. Chang, Y.-Y. Lin; Simulation of the Drusen Formation in Rpe Cells. Invest. Ophthalmol. Vis. Sci. 2010;51(13):530.
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Accumulation of lipofuscin in the RPE cells plays a major role of the drusen formation in retina. Simulation of the process of the drusen formation in RPE cells was carried out and evidenced by confocal microscopy and flow cytometry.
Photoreceptors’ outer segment (POS) were extracted from porcine retinas and fed to cultured ARPE19 cell line by the following conditions: (1) 3mg/ml POS + ARPE19 for 1, 3, and 5 hrs without additional treatment. After that the remaining POS was washed out right away; (2) ARPE19 treated by H2O2 with concentration of 0.1, 0.5 and 1.0 mM for 30 min (washed out) + 3ug/ml POS for 12 hrs, then the remaining POS was washed out immediately and continuously incubated for 0, 12 and 24 hrs. Observation of condition (1) was carried out by confocal microscopy (excitation 488 nm, emission bandpass filter 565-585 nm) and condition (2) was performed by flow cytometry.
In condition (1) it shows that the distribution of fluorescence (or fluorescent matter) was mostly surrounded the cell nucleus and the intensity was at peak which occurred at hr-3 then faded at hr-5. For pure cultured ARPE19 without feeding POS, no fluorescence is observed. In condition (2) which was performed by flow cytometry, it shows that the fluorescent intensity is largest at hr-24, then hr-12 and the hr-0 is the lowest. The ordering track is the same in each H2O2 concentration for cell treatment.The morphology of the fragment of the ‘extracted POS’ was confirmed as POS by TEM.<br
This pilot study indicates that the model of drusen formation could be manipulated by feeding POS to RPE cell line with given variable conditions.
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