METHOD OF PRODUCING TRANSPLANT FOR TREATING LIMBAL INSUFFICIENCY Russian patent published in 2020 - IPC A61F9/07 C12N5/02 

FIELD: biotechnology.

SUBSTANCE: invention refers to biotechnology, namely to obtaining a transplant for treating limbal insufficiency. Method involves mechanical cleaning of allogenic sclera, its soaking in 6 % hydrogen peroxide solution and maintenance for 4 hours (destruction of tissue pigments, lipid structures and disinfection), then washed with sterile distilled water and frozen at a temperature of -20 °C for 2 hours (which enables to destroy cell membranes). Then defrostation is soaked in water solution of ammonium hydroxide 10 % for 2 hours at temperature of 40 °C (which promotes washing out of lipids, but does not cause collagen denaturation). Then one performs washing with sterile physiological solution, soaking in 6 % hydrogen peroxide solution and holding for 6 hours. Then one performs washing with sterile normal saline, dividing into fragments 2 mm wide and 4 mm long and soaked in physiologic saline for 20 minutes. Fragments are placed in culture plates with 10 ml of culture medium DMEM/F12 with addition of 15 % of embryonal serum and added with 1 ml of concentrated cell suspension MMSC and progenitor cells of epithelium of cornea (0.5–0.6 × 10⁶ cells/ml) and incubated for 5 days.

EFFECT: disclosed is a method for producing a transplant for treating limbal insufficiency.

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