METHOD OF PRODUCING ORGAN CULTURE CHOROIDAL-PIGMENT COMPLEX FROM ADULT CADAVER DONOR'S EYE

摘要

FIELD: medicine.;SUBSTANCE: after removal of eyeball adult cadaver donor's corneal-scleral disk eyeball with no through damages of iris, ciliary body and visible part of choroid (SSO) and vitreous body outlet beyond vitreal cavity is fully submerged into a sterile container with sterile phosphate-saline buffer with pH 7.4. Complete separation of sclera from choroid by dissecting sclera meridional cuts backwards on 2, 3 or 4 "petals". On side of suprachoroidal space vorticose veins are transected and intrascleral part of optic nerve. Producing three-section choroidal pigment complex (CPC), consisting of SSO and retinal pigment epithelium (RPE). First circular incision is performed at a distance of 1 mm from a dentate line, one meridian incision in any meridian in direction from first incision backwards to an optic disk stump and second circular incision at a distance of 1 mm from optic disk stump. Further CPC carefully separated from neural retinal forceps and placed on bottom of sterile petri dishes at a position upward RPE cells. Nutrient medium is added with following composition: Dulbecco modified Eagle's medium with ham's medium F12 (DMEM/F12) - 89%, foetal calf serum - 10%, mixed antibiotics, including penicillin 10000 IU/ml, streptomycin 10000 mcg/ml, amphotericin 25 mcg/ml - 1%. Culture is incubated at 37°C and at 5% concentration CO₂, nutrient medium is replaced 2 times a day.;EFFECT: invention reduces degree of contamination of product.;1 cl, 2 dwg, 1 ex