Isolation of IPE and RPE cells: A Technique to Obtain Pigmented Epithelial Cells from Isolated Porcine Eye
Transcrição
The porcine eye contains a population of cuboidal epithelial cells that contain numerous melanosomes – organelles that help in light absorption – resulting in the pigmented appearance of these cells. Depending upon the location of these cells, epithelial cells present beneath the retina are called retinal pigment epithelial cells, or RPE cells, and the ones lining the iris are known as iris pigment epithelial cells, or IPE cells.
To isolate RPE and IPE cells, begin by taking a porcine eye. Submerge the eye in a suitable disinfectant solution to remove any microbes stuck to the eye's surface.
Remove the eye from the disinfectant solution and use a needle to make a small incision near the iris. Use this opening to make a complete circumferential cut along the iris boundary. This detaches the iris-containing anterior segment of the eye from the posterior eye bulb, which has the retina. Remove the excess portion and transfer the iris and the eye bulb into separate culture dishes.
Add a suitable culture media into the plate containing iris, and in the eye bulb. Scrape the tissue. The mechanical force loosens the epithelial cells, releasing them into the media. Aspirate the detached IPE and RPE cells. Seed them into separate culture plates and incubate till further use.
