METHOD FOR PRODUCING ORGAN CULTURE CHOROID FROM THE EYES OF AN ADULT DONOR-CORPSE Russian patent published in 2016 - IPC A61F9/07 

FIELD: medicine.

SUBSTANCE: invention relates to ophthalmology and transplantation, and relates to the preparation of organ culture choroid (OCC). To do this, the eyeball is removed from an adult donor cadaver corneal-scleral disk. Then the eyeball inspect for the presence of lesions through the iris, ciliary body and the visible part of the OCC and exit vitreous beyond vitreous cavity. In the absence of these signs eyeball completely immersed in a sterile container with sterile phosphate buffered saline with pH 7.4. Produce a complete separation of the sclera of the OCC by dissection of the scleral meridian section from front to back on the 2, 3 or 4 "petals" and the crossing from the suprachoroidal space vortice veins and intrasclerum of the optic nerve. Produces three-section choroidal pigment complex (CPC), consisting of the OCC and the retinal pigment epithelium (RPE). First circular incision is made at a distance of 1 mm from the dentate line a meridional - either in the meridian direction from the first circumferential section from front to back to the stump of the optic nerve. Another circular incision is carried out at a distance of 1 mm from the optic nerve stump. Further CPC carefully separated from the neural retinal forceps and placed on the bottom of sterile petri dishes at a position upward RPE cells. CPC is poured with 2 ml of a solution of 0.25% trypsin solution and the version in the ratio 1:1 by volume. Incubate at 37°C and 5% of CO₂ for 20 minutes. Thereafter RP is separated from the surface of OCC jet PBS pH = 7.4. OCC is transferred into a separate petri dish and washed several times with a sterile solution of phosphate buffered saline pH 7.4. By selection OCC is added the nutrient medium of the following composition: Dulbecco's modified Eagle medium with medium Ham's F12 (DMEM / F12) - 89% fetal bovine serum - 10%, the mixture of antibiotics, including penicillin 10,000 IU / ml, streptomycin 10,000 mcg / ml Amphotericin B 25 mg/ml - 1%. OCC is incubated at 37°C and concentration at 5% CO₂. This medium is replaced two times per day.

EFFECT: method reduces the degree of contamination by unauthorized cellular elements received organ culture MTR.

1 cl, 2 dwg