METHOD FOR OBTAINING ORGAN CULTURE OF A TRABECULAR NETWORK FROM A HUMAN CADAVER EYE Russian patent published in 2018 - IPC A61F9/07 C12N5/00 C12N5/07 C12N5/71 

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to ophthalmology and transplantology. To obtain the organ culture of the trabecular network from the human cadaveric eye, a circular incision of the sclera and the vascular membrane is performed, separation of the anterior pole of the eye (PSG), removal of the lens, and the separation of the PSG into parts. Donor cadaverous eye is fixed in the eyeball holder until a tonus of the eye of a living person is reached. Produce a scarification of the epithelium from the surface of the cornea and completely excise the conjunctival tissue. Before the circular incision of the sclera and the choroid, a scleral incision is made at a distance of 2 mm from the limbus with a depth not exceeding 0.1 mm. Circular trephine with a diameter of 8 mm excised and removed the remaining layers of the cornea, excluding its transition to the sclera. Then scissors make excision of the sclera and underlying tissues according to the previously outlined incision, resulting in a sample of the trabecular network, including trabeculae tissue, iris and the transitional part of the cornea in the sclera. Place the sample on the bottom of the sterile petri dish with the corneal side down and dissect it meridionally into 4 parts. Further, by magnification of the 20X microscope, iris excision is performed in each part, leaving 2 mm of tissue at its root. Then the obtained parts of the trabecular network are placed in a solution of the culture medium of the following composition: per 10 ml of medium 8,900 mcl of DMEM/F12, 1,000 mcl of 10 % fetal bovine serum, 100 mcl of 2 mM L-glutamine, 100 mcl of an antibiotic-antimycotic solution, including 10,000 IU/ml penicillin, 10,000 mcg/ml of streptomycin, 25 mcg/ml amphotericin and cultivated at 37 °C, 5 % CO₂ content and humidity 100 %. Medium is changed every 3 days.

EFFECT: method allows to increase the reliability of the results in modeling the in vivo state for the purpose of studying in the experiment, by obtaining intact, viable trabecular tissue with preservation of intravital functions and with the least amount of surrounding tissues.

1 cl, 1 ex, 1 dwg