METHOD OF EXTRACTING MOTILE SPERM CELLS FROM EJACULATE SEDIMENT IN CRYPTOZOOSPERMIA FOR PERFORMING ICSI FERTILIZATION PROCEDURE IN IVF Russian patent published in 2024 - IPC G01N33/48 A01B5/00 

FIELD: medicine.

SUBSTANCE: invention refers to in vitro fertilization (IVF) medicine and concerns a method of sperm recovery from ejaculate in cryptozoospermia for ICSI fertilization in IVF. Sperm cells are separated from other ejaculate elements by two centrifugation cycles with sampling for each subsequent centrifugation cycle, obtained as a result of the previous centrifugation cycle of the supernatant. Ejaculate treatment is carried out in the following stages: the sample is liquefied for 20-30 minutes at room temperature; whole volume of ejaculate is placed in a test tube with a conical bottom and mixed with Ooklin culture medium at ratio of 1:3; then the mixture is centrifuged for 10 minutes at 300g; supernatant is removed; obtained precipitate is resuspended with Ooklin medium to 3 ml; after which 30 mcl of Pentoxifylline is added to the suspension; then the mixture is centrifuged again for 3 minutes at 300g; supernatant is removed again; a drop is formed in a Petri dish from a nutrient medium under a layer of mineral oil at temperature of 37 °C for 10-20 minutes, wherein from the drop channels are formed, directed from the centre to the periphery; obtained precipitate is placed in the central part of the drop; in the formed thin layers of the nutrient medium channels, motile spermatozoa not contaminated with debris are visualized; obtained sperm cells are analysed by morphology for possibility of fertilization; sperm cells suitable for fertilization are caught with a needle and then used for ICSI procedure.

EFFECT: invention provides: possibility to specify diagnosis by spermogram; determine the possibility of sperm cell extraction with different degree of mobility for fertilization in the IVF/ICSI program; obtaining an aliquot of uncontaminated, debris-free sperm cells; prevents clogging of microtool; reduces the probability of bacterial introduction into the culture of human embryos; possibility of cryopreservation of the minimum required amount of sperm cells for subsequent IVF/ICSI programs, possibility of avoiding surgical intervention for a man on testicles with initial diagnosis of azoospermia, in case of isolation of single sperm cells according to the declared method; no expensive equipment of the IVF laboratory is required; saving time for the ICSI fertilization procedure; enabling the possibility of combining the ICSI fertilization dish with the disclosed method of sperm cell recovery, which increases convenience of fertilization.

2 cl, 13 dwg, 1 ex