Primer informe en Uruguay de la vitrificación de embriones como alternativa real a la técnica de congelación lenta
Palabras clave:
CRIOPRESERVACIÓN, VITRIFICACIÓN
Resumen
Introducción: la vitrificación es una técnica de criopreservación de las denominadas "rápidas". En comparación con los tradicionales procesos de congelación lenta, la estrategia de vitrificación permite la eliminación total de la formación de cristales tanto intracelular como extracelularmente. Permite poner las células directamente en el crioprotector y sumergirlas en N2 líquido. Su aplicación en el mundo ha ido creciendo exponencialmente y se puede aplicar tanto a ovocitos como a embriones en cualquier estadio.
Objetivos: revisión de la técnica de vitrificación y presentación de la experiencia de dos años en el Centro Iberoamericano de Reproducción Asistida, la primera en Uruguay.
Material y método: se incluyeron 17 pacientes de las que se han vitrificado y desvitrificado embriones. Se usó el método comercial Cryotop con las soluciones de vitrificación de Kitazato.
Resultados: con una media de embriones vitrificados de 2,71 (±0,69) por paciente, la tasa de supervivencia global de los embriones tras la desvitrificación fue de 93,4%. Con una media de embriones transferidos de 2,41 (±0,62), la tasa final de embarazo fue de 47,1%. La edad de las pacientes en las que se logra embarazo es menor (media 30,0 ±2,51) que las que no lo logran.
Conclusiones: la vitrificación como método de criopreservación se muestra prometedor y como buen sustituto de las técnicas de congelación lentas, debido a su alta tasa de éxito, su fiabilidad y la rapidez de ejecución, con resultados reproducibles.
Citas
(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.
Publicado
2010-09-30
Cómo citar
1.
Vaamonde D, Suárez Serra R, Ahumada A, Duarte C, Torres C, Oehninger S. Primer informe en Uruguay de la vitrificación de embriones como alternativa real a la técnica de congelación lenta. Rev. Méd. Urug. [Internet]. 30 de septiembre de 2010 [citado 18 de diciembre de 2024];26(3):178-86. Disponible en: https://www2.rmu.org.uy/ojsrmu311/index.php/rmu/article/view/424
Sección
Actualización de Técnicas
