Isolation of IPE and RPE cells: A Technique to Obtain Pigmented Epithelial Cells from Isolated Porcine Eye
Transcript
Clean remaining muscle tissue and skin from the eyes, and disinfect the eyes in iodine-based solution, and rinse them with PBS.
To isolate IPE cells, remove the lens and pull out the iris, as demonstrated. After preparing two irises, add 1 milliliter of complete medium and isolate the cells by scratching with a flat fire-polished Pasteur pipette, then count and seed the cells. Place the plate in the incubator.
To isolate RPE cells, remove the vitreous and retina from the posterior segment, and place the bulb in a Petri dish. Fill the bulb with 1 milliliter complete medium. Using a curved fire-polished Pasteur pipette, remove the RPE cells, collect the cell suspension within the bulb using a 1,000-microliter pipette, and transfer it into a 1.5-milliliter tube for resuspension. Then, count, and seed the cells, and incubate the plate as previously demonstrated.
