IVF 1 medium "Spermoprep" is developed for washing and isolation of motile spermatozoids from native sperm by flotation. It can be used as medium for storing viable spermatozoids before insemination and as medium for micromanipulations with spermatozoids in the ICSI procedure.
IVF 1 "Spermoprep" medium is used in assisted reproductive technologies, is intended for use in medical and preventive and research institutions.
MAIN CHARACTERISTICS:
EVF 1 "Spermoprep" media composition includes: purified water, Earl salts, glucose, lactate, pyruvate. IVF 1medium "Spermoprep" is prepared on the bicarbonate buffer basis. Since the medium provides for work both in conditions of 5-6% CO2 and in the open air, it is stabilized by HEPES. The composition of the medium includes human serum albumin tested for the presence of hepatitis B surface antigen, for antibodies to hepatitis C viruses, HIV-1 HIV-2, for antibodies to the causative agent of syphilis. IVF 1 "Spermoprep" medium can be prepared both with antibiotics and phenol red, and without these components (at customer’s request).
IVF 1 "Spermoprep" medium is sterilized by filtration through filters with pore size of 0.22 microns.
APPLICATION PROTOCOL
- Before use, the slightly opened bottle with the medium is kept in atmosphere of 5-6% CO2 at temperature of 37 ° C for 2 hours.
- After collecting the sperm, the samples are kept for 20 minutes at room temperature, then they are thoroughly mixed. If the sample is too viscous, then it is allowed to pass through a pipette with a narrow tip or through a syringe with a needle inserted, or mixing with IVF 1 "Spermoprep" in a ratio of 1:1.
- After mixing, concentration and viability of spermatozoids are evaluated under a microscope.
- Transfer 0.5 ml of sperm into a test tube, then carefully layer 1-2 ml of IVF 1 "Spermoprep" medium, aged in an incubator.
- Test tubes should be installed at an angle of 45 ° in order to increase the contact surface of the sperm with the medium. The test tubes are placed in a CO2 incubator at temperature of 37 ° C for 60 minutes. At this time, the movement of active viable spermatozoids into the medium takes place. The movement of spermatozoids into the medium can also occur at room temperature, in which case the tube lids should be tightened to prevent pH changes.
- After incubation, 0.5 ml of the supernatant is taken and concentration and viability of spermatozoids are evaluated. If the concentration of viable spermatozoids is too low, then the next 0.5 ml is taken and included in the sample.
- If the sperm concentration is normal, then 5 ml of IVF 1 "Spermoprep" medium is added to the sample, mixed well, centrifuged at 1500 rpm for 15 minutes.
- The supernatant is removed, the precipitate is resuspended in an equal volume of the balanced IVF 1 "Spermoprep" medium.
- The prepared sperm sample is wrapped in aluminum foil and stored at room temperature (22 ° C) until insemination, approximately 6 hours after oocyte collection.
SHELF LIFE, STORAGE AND TRANSPORTATION CONDITIONS
Shelf life is 3 months from the date of manufacture. Overdue nutrient medium is not subject to use.
Store and transport at temperature of 2 to 8 ° C in a place protected from light in accordance with SR 3.3.2.1248-03. In case of non-compliance with the storage and transportation conditions, the medium is subject to disposal.
Do not freeze!
Unused residue of the heated medium is not subject to reheating and use.
The product must be used within 7 days from the bottle first opening.
Open the vial in sterile conditions.
