IVF 1 "Universal" medium with antibiotics with phenol red

IVF 1 "Universal" medium is developed for fertilization. It can be used for short-term cultivation of embryos at an early stage of development. The medium is also suitable for ICSI.

IVF 1 "Universal" medium is used in assisted reproductive technologies, is intended for use in medical and preventive and research institutions.

This product has Registration Certificate.

THE MAIN CHARACTERISTICS:

IVF 1 "Universal" medium is prepared on bicarbonate buffer basis. Composition: purified water, Earl salts, glucose, lactate, pyruvate, amino acids, human serum albumin. Albumin was 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 "Universal" medium can be prepared with the addition of antibiotics and phenol red dye, or without these components (at customer’s request). IVF 1 "Universal" medium is sterilized by filtering through filters with pore size of 0.22 microns

APPLICATION PROTOCOL

The following protocol is recommended for fertilization and short-term embryo cultivation:

1. 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.
2. For successful fertilization, it is required to use spermatozoids in optimal concentration: 80,000 -100,000 motile spermatozoids in 1 ml of medium.
3. Oocyte-Cumulus complexes after washing in  IVF 1 "Oaklyn" medium placed in IVF 1 "Universal" medium and incubated for 3-4 h.
4. After this time the drops with the medium containing oocyte-Cumulus complexes, making the volume of washed spermatozoids up to the final concentration in the medium 80000-100000 motile sperm in 1 ml of medium.
5. Immediately after addition of spermatozoids, the tablet is transferred to a CO2 incubator.
6. After 16 -20 h, the oocytes are carefully washed, transferring through a few drops with balanced IVF 1 "Universal" medium. Then the formation of pronuclei in the zygotes is checked, after which the normally formed zygotes are carefully washed and transferred to an embryo culture medium (IVF PRO "Droblenie" or into a new portion of IVF medium 1 "Universal").
7. Cultivation in IVF 1 "Universal" medium can be carried out up to the stage of two blastomeres, then the embryos should be placed in the medium for subsequent cultivation up to the stage of 4-8 blastomeres (IVF PRO"Droblenie").
8. Fertilization and cultivation can be carried out in microdrops of the medium coated with liquid paraffin oil, or in wells with a medium volume of 500 µl without coating the medium with oil.

To use the medium during ICSI, the following protocol is recommended:

1. Before use, the slightly opened bottle with the medium should be kept in atmosphere of 5-6% CO2 at temperature of 37 ° C for 2 hours.
2. Oocyte-cumulus complexes after washing in the  IVF 1 medium "Ooklin" are placed in the of IVF 1 "Universal" medium. Incubation in IVF 1 "Universal" medium should be carried out for 3-4 hours.
3. Then, the release of oocytes from cumulus cells is carried out by means of short-term exposure in IVF 1 medium "GIAZA" and subsequent denudation of oocytes in IVF 1 medium "Universal".
4. After denudation and before sperm injection, oocytes should be kept in a CO2 incubator in IVF 1 "Universal" medium for a short period of time.
5. The procedure of sperm injection is carried out in Petri dishes.  A drop of IVF 1 "PVP" medium is added into the middle of the cup. IVF 1 "Universal" medium drops are placed into the cup, where injections will be carried out. The number of these drops corresponds to the number of oocytes.
6. All drops should be covered with a layer of liquid paraffin oil, previously aged in atmosphere of 5-6% CO2 at temperature of 37 ° C for 12 hours.
7. It is recommended to leave the cup with oil-covered drops of the medium in the CO2 incubator for a while so that the medium pH and the temperature return to the required values.
8. The prepared spermatozoids are placed in the central drop. Oocytes are added into peripheral drops.
9. One sperm is fixed with use of injection micropipette. Sperm injection is recommended to be carried out in a minimum volume of IVF 1medium "PVP".
10. Then the oocyte is fixed with use of a holding pipette. It is important to fix the oocyte in such a way that the polar body is in position for 6 or 12 hours.
11. An injectable micropipette is injected into the oocyte at position of 3 h. To confirm the puncture of the shiny shell, it is recommended to aspirate part of the cytoplasm.
12. If the pipette has penetrated into the cytoplasm, then the sperm with part of the aspirated cytoplasm is injected into the oocyte.
13. After the sperm addition, pressure is applied in the holding pipette and the oocyte is disconnected.
14. After the injection, oocytes are washed and incubated in a balanced medium of IVF PRO "Droblenie" or IVF 1 "Universal".
15. From the moment of injection, 16-18 hours should pass, after which the formation of pronuclei in oocytes is checked. Further cultivation is carried out in the same media.
16. Cultivation can be carried out up to the stage of two blastomeres in the IVF 1 "Universal" medium, then the embryos should be placed in IVF PRO "Droblenie" medium for subsequent cultivation up to the stage of 4-8 blastomeres, or transferred to the uterine cavity for implantation in IVF PRO "Shvartovaya" medium.

SHELF LIFE, STORAGE AND TRANSPORTATION CONDITIONS

Shelf life is 3 months from the date of manufacture, subject to the storage regime.

Store and transport in accordance with SR 3.3.2.1248-03 at temperature of 2 to 8 ° C in a place protected from light.

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.

It is recommended to store the medium with a minimum volume of air in the bottle.

Open the vial in sterile conditions.