METHOD OF OBTAINING SUSPENSION OF HUMAN CORNEAL ENDOTHELIAL CELLS USING ENZYME 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 (CD) is formed from it under sterile conditions using a trephine, it is placed on a sterile surface with the endothelium facing up, then the edges of the Descemet membrane (DM) are separated from the anterior ring of Schwalbe over 180 degrees. Using tweezers, the DM is peeled off towards the center by half its area, folding the posterior epithelium towards each other. Using a trepanation blade, an incision is made from the posterior stroma of the cornea, the DM with endothelium is separated with tweezers and transferred to an Eppendorf tube, 0.3 ml of the recombinant TrypLE enzyme is added, then placed in a thermal shaker for 5 minutes, at 300 rpm at 37 degrees Celsius. Next, the contents of the tube are inactivated by 10-fold dilution using a solution for storing the cornea, followed by centrifugation for 5 minutes, at 900 rpm at 37 degrees Celsius.

EFFECT: method makes it possible to obtain a suspension of endothelial cells from the human cadaveric eyeball, which is an accessible and easily reproducible procedure in the presence of standard laboratory equipment, which simplifies the possibilities of its use.

1 cl, 1 ex