First report in Uruguay on vitrification of embryos as a real alternative to slow freezing
Keywords:
CRYOPRESERVATION, VITRIFICATIONAbstract
Introduction: vitrification is a cryopreservation technique that forms part of the so called "fast" cryopreservation techniques. Compared to traditional slow freezing processes, vitrification enables the total elimination of both intracellular and extracellular crystal formation.
It allows for placing the cells directly on the cryoprotector and submersing them in liquid nitrogen. Its application around the world has exponentially grown and the technique may be applied both to ovocytes and embryos at all stages.
Objectives: to carry out a review of the vitrification technique, the first one to be conducted in Uruguay, and to present the two-year experience at the Ibero-American Center for Assisted Reproduction.
Method: we included 17 patients in our study, the embryos of which have been vitrified and unvitrified. We used the commercial Cryotop method with the Kitazato vitrification solutions.
Results: with an average of 2,71 (±0,69) vitrified embryos per patients, global embryo survival rate after vitrification was 93,4%. With an average of 2,41 (±0,62) embryos transferred , the final pregnancy rate was 47,1%. The average age of patients who succeeded in getting pregnant is lower (average 30,0 ±2,51) than those who fail to get pregnant.
Conclusions: vitrification as a cryopreservation method appears to be a promising technique and seems to be a good substitute for show freezing technique, due to its high-success rates, its reliability and execution speed, with replicable results.
References
(1) De Mouzon J. Session 50 – IVF Monitoring in Europe and Worldwide Hum Reprod 2006; 21(suppl 1): i76.
(2) Liebermann J. Vitrification of human blastocysts: an update. Reprod Biomed Online 2009; 19(Suppl 4): 105-14.
(3) Rall WF, Fahy GM. Ice-free cryopreservation of mouse embryos at -196 degrees C by vitrification. Nature 1985; 313(6003): 573-5.
(4) Trounson A, Mohr L. Human pregnancy following cryopreservation, thawing and transfer of an eight-cell embryo. Nature 1983; 305(5936): 707-9.
(5) Vajta G, Kuwayama M. Improving cryopreservation systems. Theriogenology 2006; 65(1): 236-44.
(6) Fahy GM, Wowk B, Wu J, Paynter S. Improved vitrification solutions based on the predictability of vitrification solution toxicity. Cryobiology 2004; 48(1): 22-35.
(7) Mazur P. Freezing of living cells: mechanisms and implications. Am J Physiol 1984; 247(3 Pt1): C125-42.
(8) Luyet B, Hodapp A. Revival of frog’s spermatozoa vitrified in liquid air. Proc Meet Soc Exp Biol 1938; 39: 433-4.
(9) Fahy GM, MacFarlane DR, Angell CA, Meryman HT. Vitrification as an approach to cryopreservation. Cryobiology 1984; 21(4): 407-26.
(10) Fahy GM. Vitrification: a new approach to organ cryopreservation. Prog Clin Biol Res 1986; 224: 305-35.
(11) Rall WF. Factors affecting the survival of mouse embryos cryopreserved by vitrification. Cryobiology 1987; 24(5): 387-402.
(12) Arav A, Zeron Y, Leslie SB, Behboodi E, Anderson GB, Crowe JH. Phase transition temperature and chilling sensitivity of bovine oocytes. Cryobiology 1996; 33(6): 589-99.
(13) Ghetler Y, Yavin S, Shalgi R, Arav A. The effect of chilling on membrane lipid phase transition in human oocytes and zygotes. Hum Reprod 2005; 20(12): 3385-9.
(14) Kasai M, Mukaida T. Cryopreservation of animal and human embryos by vitrification. Reprod BioMed Online 2004; 9(2): 164-70.
(15) Kasai M, Komi JH, Takakamo A, Tsudera H, Sakurai T, Machida T. A simple method for mouse embryo cryopreservation in a low toxicity vitrification solution, without appreciable loss of viability. J Reprod Fertil 1990; 89(1): 91-7.
(16) Emiliani S, Van den Bergh M, Vannin AS, Biramane J, Englert Y. Comparison of ethylene glycol, 1,2-propanediol and glycerol for cryopreservation of slow-cooled mouse zygotes, 4-cell embryos and blastocysts. Hum Reprod 2000; 15(4): 905-10.
(17) Zhu SE, Kasai M, Otoge H, Sakurai T, Machida T. Cryopreservation of expanded mouse blastocysts by vitrification in ethylene glycol-based solutions. J Reprod Fertil 1993; 98(1): 139-45.
(18) Yokota Y, Sato S, Yokota M, Ishikawa Y, Makita M, Asada T, et al. Successful pregnancy following blastocyst vitrification. Hum Reprod 2000; 15(8): 1802-3.
(19) Critser JK, Agca Y, Gunasena KT. The cryobiology of mammalian oocytes. In Karow AM, Critser JK, eds. Reproductive Tissue Banking: scientific principles. New York: Academic Press, 1997: 329-57.
(20) Kuleshova LL, MacFarlane DR, Trounson AO, Shaw JM. Sugars exert a major influence on the vitrification properties of ethylene glycol-based solutions and have a low toxicity to embryos and oocytes. Cryobiology 1999; 38(2): 119-30.
(21) Kuleshova LL, Shaw JM, Trounson AO. Studies on replacing most of the penetrating cryoprotectant by polymers for embryo cryopreservation. Cryobiology 2001; 43(1): 21-31.
(22) Chen C. Pregnancy after human oocyte cryopreservation. Lancet 1986; 1(8486): 884-6.
(23) van Uem JF, Siebzehnrübl ER, Schuh B, Koch R, Trotnow S, Lang N. Birth after cryopreservation of unfertilized oocytes. Lancet 1987; 1(8535): 752-3.
(24) Tucker MJ, Wright G, Morton PC, Massey JB. Birth after cryopreservation of immature oocytes with subsequent in vitro maturation. Fertil Steril 1998; 70(3): 578-9.
(25) Porcu E, Fabbri R, Seracchioli R, Ciotti PM, Magrini O, Flamigni C. Birth of a healthy female after intracytoplasmic sperm injection of cryopreserved human oocytes. Fertil Steril 1997; 68(4): 724-6.
(26) Ruffing NA, Steponkus PL, Pitt RE, Parks JE. Osmometric behavior, hydraulic conductivity, and incidence of intracellular ice formation in bovine oocytes at different developmental stages. Cryobiology 1993; 30(6): 562-80.
(27) Pedro PB, Zhu SE, Makino N, Sakurai T, Edashige K, Kasai M. Effects of hypotonic stress on the survival of mouse oocytes and embryos at various stages. Cryobiology 1997; 35(2): 150-8.
(28) Hunter JE, Fuller BJ, Bernard A, Jackson A, Shaw RW. Vitrification of human oocytes following minimal exposure to cryoprotectants; initial studies on fertilization and embryonic development. Hum Reprod 1995; 10(5): 1184-8.
(29) Liebermann J, Tucker MJ. Comparison of vitrification and conventional cryopreservation of day 5 and day 6 blastocysts during clinical application. Fertil Steril 2006; 86(1): 20-6.
(30) Osada H, Aono F, Kuwayama M, Morita H, Teramoto S, Kato O. Clinical efficiency of vitrification on blastocysts transfer cycles. Fertil Steril 2003; 80(Suppl 3): S63.
(31) Vanderzwalmen P, Bertin G, Debauche Ch, Standaert V, Bollen N, van Roosendaal E, et al. Vitrification of human blastocysts with the Hemi-Straw carrier: application of assisted hatching after thawing. Hum Reprod 2003; 18(7): 1504-11.
(32) Kuwayama M, Vajta G, Kato O, Leibo SP. Highly efficient vitrification method for cryopreservation of human oocytes. Reprod BioMed Online 2005; 11(3): 300-8.
(33) Kuwayama M, Vajta G, Ieda S, Kato O. Comparison of open and closed methods for vitrification of human embryos and the elimination of potential contamination. Reprod BioMed Online 2005; 11(): 608-14.
(34) Vajta G, Holm P, Kuwayama M, Booth PJ, Jacobsen H, Greve T, et al. Open pulled straw (OPS) vitrification: a new way to reduce cryoinjuries of bovine ova and embryos. Mol Reprod Dev 1998; 51(1): 53-8.
(35) Ishimori H, Saeki K, Inai M, Nagao Y, Itasaka J, Miki Y, et al. Vitrification of bovine embryos in a mixture of ethylene glycol and dimethyl sulfoxide. Theriogenology 1993; 40(2): 427-33.
(36) Isachenko V, Montag M, Isachenko E, Dessole S, Nawroth F, van der Ven H. Aseptic vitrification of human germinal vesicle oocytes using dimethyl sulfoxide as a cryoprotectant. Fertil Steril 2006; 85(3): 741-7.
(37) Vajta G, Nagy ZP. Are programmable freezers still needed in the embryo laboratory? Review on vitrification. Reprod BioMed Online 2006; 12(6): 779-96.
(38) Michelmann HW, Nayudu P. Cryopreservation of human embryos. Cell Tissue Bank 2006; 7(2): 135-41.
(39) Kuwayama M. Highly efficient vitrification for cryopreservation of human oocytes and embryos: the Cryotop method. Theriogenology 2007; 67(1): 73-80.
(40) Bielanski A, Vajta G. Risk of contamination of germplasm during cryopreservation and cryobanking in IVF units. Hum Reprod 2009; 24(10): 2457-67.
(2) Liebermann J. Vitrification of human blastocysts: an update. Reprod Biomed Online 2009; 19(Suppl 4): 105-14.
(3) Rall WF, Fahy GM. Ice-free cryopreservation of mouse embryos at -196 degrees C by vitrification. Nature 1985; 313(6003): 573-5.
(4) Trounson A, Mohr L. Human pregnancy following cryopreservation, thawing and transfer of an eight-cell embryo. Nature 1983; 305(5936): 707-9.
(5) Vajta G, Kuwayama M. Improving cryopreservation systems. Theriogenology 2006; 65(1): 236-44.
(6) Fahy GM, Wowk B, Wu J, Paynter S. Improved vitrification solutions based on the predictability of vitrification solution toxicity. Cryobiology 2004; 48(1): 22-35.
(7) Mazur P. Freezing of living cells: mechanisms and implications. Am J Physiol 1984; 247(3 Pt1): C125-42.
(8) Luyet B, Hodapp A. Revival of frog’s spermatozoa vitrified in liquid air. Proc Meet Soc Exp Biol 1938; 39: 433-4.
(9) Fahy GM, MacFarlane DR, Angell CA, Meryman HT. Vitrification as an approach to cryopreservation. Cryobiology 1984; 21(4): 407-26.
(10) Fahy GM. Vitrification: a new approach to organ cryopreservation. Prog Clin Biol Res 1986; 224: 305-35.
(11) Rall WF. Factors affecting the survival of mouse embryos cryopreserved by vitrification. Cryobiology 1987; 24(5): 387-402.
(12) Arav A, Zeron Y, Leslie SB, Behboodi E, Anderson GB, Crowe JH. Phase transition temperature and chilling sensitivity of bovine oocytes. Cryobiology 1996; 33(6): 589-99.
(13) Ghetler Y, Yavin S, Shalgi R, Arav A. The effect of chilling on membrane lipid phase transition in human oocytes and zygotes. Hum Reprod 2005; 20(12): 3385-9.
(14) Kasai M, Mukaida T. Cryopreservation of animal and human embryos by vitrification. Reprod BioMed Online 2004; 9(2): 164-70.
(15) Kasai M, Komi JH, Takakamo A, Tsudera H, Sakurai T, Machida T. A simple method for mouse embryo cryopreservation in a low toxicity vitrification solution, without appreciable loss of viability. J Reprod Fertil 1990; 89(1): 91-7.
(16) Emiliani S, Van den Bergh M, Vannin AS, Biramane J, Englert Y. Comparison of ethylene glycol, 1,2-propanediol and glycerol for cryopreservation of slow-cooled mouse zygotes, 4-cell embryos and blastocysts. Hum Reprod 2000; 15(4): 905-10.
(17) Zhu SE, Kasai M, Otoge H, Sakurai T, Machida T. Cryopreservation of expanded mouse blastocysts by vitrification in ethylene glycol-based solutions. J Reprod Fertil 1993; 98(1): 139-45.
(18) Yokota Y, Sato S, Yokota M, Ishikawa Y, Makita M, Asada T, et al. Successful pregnancy following blastocyst vitrification. Hum Reprod 2000; 15(8): 1802-3.
(19) Critser JK, Agca Y, Gunasena KT. The cryobiology of mammalian oocytes. In Karow AM, Critser JK, eds. Reproductive Tissue Banking: scientific principles. New York: Academic Press, 1997: 329-57.
(20) Kuleshova LL, MacFarlane DR, Trounson AO, Shaw JM. Sugars exert a major influence on the vitrification properties of ethylene glycol-based solutions and have a low toxicity to embryos and oocytes. Cryobiology 1999; 38(2): 119-30.
(21) Kuleshova LL, Shaw JM, Trounson AO. Studies on replacing most of the penetrating cryoprotectant by polymers for embryo cryopreservation. Cryobiology 2001; 43(1): 21-31.
(22) Chen C. Pregnancy after human oocyte cryopreservation. Lancet 1986; 1(8486): 884-6.
(23) van Uem JF, Siebzehnrübl ER, Schuh B, Koch R, Trotnow S, Lang N. Birth after cryopreservation of unfertilized oocytes. Lancet 1987; 1(8535): 752-3.
(24) Tucker MJ, Wright G, Morton PC, Massey JB. Birth after cryopreservation of immature oocytes with subsequent in vitro maturation. Fertil Steril 1998; 70(3): 578-9.
(25) Porcu E, Fabbri R, Seracchioli R, Ciotti PM, Magrini O, Flamigni C. Birth of a healthy female after intracytoplasmic sperm injection of cryopreserved human oocytes. Fertil Steril 1997; 68(4): 724-6.
(26) Ruffing NA, Steponkus PL, Pitt RE, Parks JE. Osmometric behavior, hydraulic conductivity, and incidence of intracellular ice formation in bovine oocytes at different developmental stages. Cryobiology 1993; 30(6): 562-80.
(27) Pedro PB, Zhu SE, Makino N, Sakurai T, Edashige K, Kasai M. Effects of hypotonic stress on the survival of mouse oocytes and embryos at various stages. Cryobiology 1997; 35(2): 150-8.
(28) Hunter JE, Fuller BJ, Bernard A, Jackson A, Shaw RW. Vitrification of human oocytes following minimal exposure to cryoprotectants; initial studies on fertilization and embryonic development. Hum Reprod 1995; 10(5): 1184-8.
(29) Liebermann J, Tucker MJ. Comparison of vitrification and conventional cryopreservation of day 5 and day 6 blastocysts during clinical application. Fertil Steril 2006; 86(1): 20-6.
(30) Osada H, Aono F, Kuwayama M, Morita H, Teramoto S, Kato O. Clinical efficiency of vitrification on blastocysts transfer cycles. Fertil Steril 2003; 80(Suppl 3): S63.
(31) Vanderzwalmen P, Bertin G, Debauche Ch, Standaert V, Bollen N, van Roosendaal E, et al. Vitrification of human blastocysts with the Hemi-Straw carrier: application of assisted hatching after thawing. Hum Reprod 2003; 18(7): 1504-11.
(32) Kuwayama M, Vajta G, Kato O, Leibo SP. Highly efficient vitrification method for cryopreservation of human oocytes. Reprod BioMed Online 2005; 11(3): 300-8.
(33) Kuwayama M, Vajta G, Ieda S, Kato O. Comparison of open and closed methods for vitrification of human embryos and the elimination of potential contamination. Reprod BioMed Online 2005; 11(): 608-14.
(34) Vajta G, Holm P, Kuwayama M, Booth PJ, Jacobsen H, Greve T, et al. Open pulled straw (OPS) vitrification: a new way to reduce cryoinjuries of bovine ova and embryos. Mol Reprod Dev 1998; 51(1): 53-8.
(35) Ishimori H, Saeki K, Inai M, Nagao Y, Itasaka J, Miki Y, et al. Vitrification of bovine embryos in a mixture of ethylene glycol and dimethyl sulfoxide. Theriogenology 1993; 40(2): 427-33.
(36) Isachenko V, Montag M, Isachenko E, Dessole S, Nawroth F, van der Ven H. Aseptic vitrification of human germinal vesicle oocytes using dimethyl sulfoxide as a cryoprotectant. Fertil Steril 2006; 85(3): 741-7.
(37) Vajta G, Nagy ZP. Are programmable freezers still needed in the embryo laboratory? Review on vitrification. Reprod BioMed Online 2006; 12(6): 779-96.
(38) Michelmann HW, Nayudu P. Cryopreservation of human embryos. Cell Tissue Bank 2006; 7(2): 135-41.
(39) Kuwayama M. Highly efficient vitrification for cryopreservation of human oocytes and embryos: the Cryotop method. Theriogenology 2007; 67(1): 73-80.
(40) Bielanski A, Vajta G. Risk of contamination of germplasm during cryopreservation and cryobanking in IVF units. Hum Reprod 2009; 24(10): 2457-67.
Downloads
Published
2010-09-30
How to Cite
1.
Vaamonde D, Suárez Serra R, Ahumada A, Duarte C, Torres C, Oehninger S. First report in Uruguay on vitrification of embryos as a real alternative to slow freezing. Rev. Méd. Urug. [Internet]. 2010 Sep. 30 [cited 2024 Nov. 21];26(3):178-86. Available from: https://revista.rmu.org.uy/index.php/rmu/article/view/424
Issue
Section
Techniques Update
