METHOD FOR PRODUCING DEDIFFERENTIATED CELLS OF ADULT RETINAL PIGMENT EYE EPITHELIUM

摘要

FIELD: medicine.;SUBSTANCE: eyeball is enucleated in autopsy, washed in an alcoholic solution and in Ca²⁺ Mg²⁺ -free Hanks' solution with an antibiotic added; an anterior segment of an eyeball is removed along a dentate line, a vitreous body and a neutral retina are separated from a pigment epithelium; an eyecup is filled with Ca²⁺ Mg²⁺ -free Hanks' solution with EDTA, and retinal pigment epithelial cells are incubated for 15-30 min; the produced cells are pipetted and transferred to a sterile recovery medium, pipetted, centrifuged; the recovered cells are resuspended in a high-serum growth medium, then the prepared suspension of the retinal pigment epithelial cells is distributed over a culture surfaces and cultivated until the attached cells reach 15-25 % of its area; the suspension with the unattached cells is aspirated, distributed over the fresh culture surfaces and cultivated; and the unattached cells are added with the high-serum growth medium and cultivated until a confluent finish monolayer is produced.;EFFECT: invention allows eliminating damaging action of enzymes and ensuring a lower adhesion cell population to attach on the culture surfaces.;13 cl, 4 dwg, 1 tbl, 3 ex