ENZYMATIC METHOD OF OBTAINING SUSPENSION OF HUMAN CORNEAL ENDOTHELIAL CELLS Russian patent published in 2023 - IPC A61B10/02 C12N5/71 A61F9/07 

FIELD: medicine; ophthalmology.

SUBSTANCE: corneal epithelium is completely removed from the surface of the human cadaver eye, and a corneoscleral disc (CSD) is formed from it under sterile conditions using a trephine. Then, using tweezers, the CSD is held by the edge of the scleral rim, placed parallel to a sterile Petri dish with the endothelial surface of the cornea upward, and a proteolytic, collagenolytic or recombinant enzyme solution with a volume of 1 ml is installed on it, exposure time is 3, 20, 15 minutes, respectively. Then, atraumatic aspiration of the resulting solution consisting of corneal endothelial cells and an enzyme solution is performed, after which the resulting solution is poured into a sterile tube, 5 ml of corneal storage solution is added and centrifugation is performed at 800 rpm for 5 minutes at a temperature of 37 degrees Celsius. After centrifugation, the supernatant is removed with an aspirator with a sterile tip, 5 ml of cornea storage solution is added to the sediment, and the resulting suspension is taken into a test tube using a mechanical dispenser with a sterile tip, which is stored at a temperature of 4 degrees Celsius.

EFFECT: method will make it possible to obtain a suspension of endothelial cells from the human cadaveric eyeball by instillation of an enzyme solution on the endothelial surface of the cornea, aspiration and centrifugation of the resulting solution.

1 cl, 3 ex