Cryotech Vitrification Kit 101
“Cryotech® Vitrification Kit 101”
At GFS, we take pride in offering world-class IVF products to all our clients and patients. We aim to serve you with the best results and experience while utilizing globally recognized vitrification techniques. Moreover, our following package of “Cryotech® Vitrification Kit 101”
Contents of Cryotech® Vitrification Kit 101”
- Equilibration Solution (ES): 1 vial of 1.0ml.
- Vitrification Solution (VS): 2 vials of 1.0ml.
- 4 Cryotecs: 1 Cryotec can contain more than one oocyte/embryo (recommend 3-4 maximum for oocytes and 4-cell embryos and one for blastocyst for single embryo transfer).
- -3 Vitri Plates with 3 wells each.
Guidelines to use Cryotech® Vitrification Kit 101”
- -Room temperature (25–27ºC) should be used for the entire process.
- -Pour nitrogen into a container.
- -Make a note of the thickness of the oocyte and compare it to the periviteline space in the zona pellucida.
- It’s crucial to use a Pasteur pipette with the appropriate diameter for both blastocysts (160-200 µm) and oocyte embryos (140-150 µm).
Equilibration of oocytes and embryos
- Transfer 300 µl of ES into each of the 1º, 2º, and 3º wells on the Vitri Plate.
- Place the oocyte/embryo in the 1½ well on the ES surface.
- The embryo or oocyte will sink and start to shrink back to its initial size (blastocysts and oocytes can take up to 15 minutes, and other stages of the embryo can take up to 12 minutes).
Please take note: You have 25 seconds to complete the following steps, with a maximum of 90 seconds.
- Use VS to transfer the oocyte or embryo to the bottom of the 2° well (Instead of starting with a minimum volume of ES.) The oocyte/embryo washes up and instantly floats to the surface of VS.
- Take just the oocyte or embryo and place it in the bottom of the well after cleaning the pipette’s interior wall with new VS media. Hold off until the embryo/oocyte stops floating in VS.
- Place the oocyte or embryo in the center of the 3° well with VS, and use a pipette to mix the media five times.
- Using the pipette, remove only the oocyte or embryo at the tip, and place it on the end of the cryotec seat with the least amount of VS.
- Drop the Cryotech into liquid nitrogen right away.
- Insert the cap and keep it in a nitrogen tank for storage.
Quality Control Tests:
This Lot Nº JIHA0115 (All Solutions)
Our Cryotech® Vitrification Kit 101” has successfully passed the following controls:
- Sterility: Sterility test.
- Endotoxin by ES methodology (Each component).
- Efficiency: survival of 50/50 Mouse embryos and Porcine oocytes.
Stability and storage
Kits and solutions should be transported at room temperature and then refrigerated between 2 and 8 degrees Celsius until their expiration date.
- -Modified HEPES Buffered MEM
- -Hydroxy Propyl Cellulose
- -Ethylene Glycol
- -Dimethyl Sulfoxide
- -Endotoxin-free Trehalose
- Kuwayama M. Highly efficient vitrification for cryopreservation of human oocytes and embryos: The CryoTop method. Theriogenology 67, 73 2007.
- Cobo A, Kuwayama M. Comparison of concomitant outcome achieved with fresh and cryopreserved donor oocytes vitrified by the Cryotop method. Fertil Steril. J89(6): 1657-64, 2007.
- Antinori M, Licata E, Dani G, Cerusico F, Versaci C, Antinori S. Cryotop vitrification of human oocytes results in a high survival rate and healthy deliveries. Reproductive BioMedicine Online 14, 5-667, 2007.
- Vajta G, Kuwayama M. Improving cryopreservation systems. Theriogenology 65(1), 236-44, 2006.
- Kuwayama M. Highly efficient vitrification method for cryopreservation of human oocytes. Reproductive BioMedicine Online 11:300-308, 2005.
- Ushijima J, Kuwayama M. High survival rate of bovine oocytes matured in vitro following vitrification. J Reprod Dev. 50:685-96, 2004.
- Fukui Y, Kuwayama M. Effect of cryo-device type and donor’s sexual maturity on vitrification of minke whale oocytes at germinal vesicle stage. Zygote 12, 333-338, 2004.
- Hochi S, Kuwayama M. Improved Survival of Vitrified in vivo-derived porcine embryos. J. Reprod. Develop. 50, 481-486, 2004.
- Esaki R, Kuwayama M. Cryopreservation of porcine embryos derived from in vitri- matured oocytes. Biology of Reproduction. 71, 432-437, 2004.
Please Note: This “Cryotech® Vitrification Kit 101” is for in vitro use only!