METHOD FOR ISOLATING SPERM FROM TESTICULAR BIOPTATE IN PATIENTS WITH DIAGNOSIS OF NON-OBSTRUCTIVE AZOOSPERMIA FOR DIAGNOSIS OF THE PRESENCE OF MATURE FORMS OF SPERMATOGENESIS AND THEIR USE IN FERTILIZATION IN THE IVF/ICSI PROGRAM IN MARITAL MARRIED COUPLES WITH DIFFERENT FORMS OF INFERTILITY Russian patent published in 2023 - IPC A61B17/435 G01N33/483 

FIELD: medicine.

SUBSTANCE: method for isolating sperm from a testicular biopsy in patients diagnosed with non-obstructive azoospermia. The method for isolating sperm from a testicular biopsy in patients diagnosed with non-obstructive azoospermia, characterized by enzymatic destruction of the biopsy, differs by the fact that on the eve of the operation, collagenase type VI (Collagen Type VI (COL6)) is weighed at a concentration of 3 mg/ml based on the volume of the biopsy obtained, weighed diluted in PBS buffer immediately upon receipt of the biopsy specimen in the laboratory, transportation is carried out in Ooklin medium, the transport medium is washed with a physical solution, the entire volume of the biopsy specimen is placed in a conical tube graduated in ml, the resulting volume is measured in a 1:1 ratio, the volume of the measured biopsy specimen is combined with the solution collagenase in PBS buffer, mixed and set for 30 minutes at 37°C into the incubator, after 30 minutes the test tube is shaken and the formation of a homogeneous suspension of cells in the solution is monitored; when a homogeneous suspension is obtained, it is immediately centrifuged for 3 minutes at 300 g, the supernatant is removed, the resulting sediment is diluted with Ooklin medium to 5 ml and thoroughly mixed, the cell suspension is centrifuged for 3 minutes at 300 g, the supernatant is removed, the sediment is diluted with 2-3 ml of Ooklin medium, mixed using dispenser pipette with a 1.0 ml spout, an aliquot of the suspension is taken for microscopy of mature forms of spermatogenesis, if spermatozoa are detected, they are photographed and assessed for the possibility of use in the IVF program, immobile spermatozoa are exposed to the drug Pentoxifylline to assess their suitability for use in the IVF program, mobile spermatozoa are frozen and/or used in the IVF/ICSI program.

EFFECT: processing the biopsy in its entire volume, thereby increasing the speed and quality of the study, reducing time costs in the IVF/ICSI program.

1 cl, 20 dwg, 2 tbl, 1 ex