Clinical and Experimental Reproductive Medicine

The Korean Society for Reproductive Medicine (대한생식의학회)
권호 표지
DOI QR코드

DOI QR Code

Vitrification solution without sucrose for cryopreservation in mouse blastocysts

  • Joo, Jong Kil (Department of Obstetrics and Gynecology, Pusan National University School of Medicine and Medical Research Institute, Pusan National University Hospital) ;
  • Lee, Young Ju (Department of Obstetrics and Gynecology, Pusan National University School of Medicine and Medical Research Institute, Pusan National University Hospital) ;
  • Jeong, Ju Eun (Department of Obstetrics and Gynecology, Pusan National University School of Medicine and Medical Research Institute, Pusan National University Hospital) ;
  • Kim, Seung Chul (Department of Obstetrics and Gynecology, Pusan National University School of Medicine and Medical Research Institute, Pusan National University Hospital) ;
  • Ko, Gyoung Rae (Infertililty Center, Pusan National University Hospital) ;
  • Lee, Kyu Sup (Department of Obstetrics and Gynecology, Pusan National University School of Medicine and Medical Research Institute, Pusan National University Hospital)
  • Received : 2014.07.24
  • Accepted : 2014.08.22
  • Published : 2014.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

Objective: This study was designed to investigate the survival rate of vitrified mouse blastocysts depending on the presence or absence of sucrose in vitrification solution. Methods: Mouse two-cell embryos were collected and cultured to blastocysts. Two vitrification solutions were prepared. The control solution was composed of 25% glycerol, 25% ethylene glycol, and 0.5 M sucrose (G25E250.5S) containing 2.5 mL glycerol, 2.5 mL ethylene glycol, 2 mL SSS, and 0.855 g sucrose in 5 mL PB1. The experimental solution was composed of 25% glycerol and 25% ethylene glycol (G25E25) and contained 2.5 mL glycerol and 2.5 mL ethylene glycol in 5 mL PB1. Artificial shrinkage was conducted by aspirating the blastocoelic fluid using an ICSI pipette. To examine the effect of sucrose in the vitrification solution on the survival rate of mouse blastocysts, the shrunken-equilibrated blastocysts were rehydrated or vitrified after being exposed to one of the two vitrification solutions. After exposure and the vitrification-thawing process, the re-expansion rate and hatching rate were evaluated after 6 hours of in vitro culture. Results: The re-expansion rate of mouse blastocysts exposed to vitrification solution with and without sucrose were not different in the experimental solution (without sucrose) (98%) and the control solution (with sucrose) (92%) (p>0.05). The hatching rate was higher in the experimental solution (95%) than in the control solution (88%), but did not differ across two treatments (p>0.05). The re-expansion rate of mouse blastocysts vitrified in the control solution was 92% and 94%, respectively (p>0.05), and the hatching rate was higher in the experimental solution (90%) than in the control solution (74%) (p<0.05). Conclusion: Sucrose need not be added in vitrification solution for freezing of artificially shrunken mouse blastocysts.

Keywords

References

  1. Gardner DK, Schoolcraft WB, Wagley L, Schlenker T, Stevens J, Hesla J. A prospective randomized trial of blastocyst culture and transfer in in-vitro fertilization. Hum Reprod 1998;13:3434-40. https://doi.org/10.1093/humrep/13.12.3434
  2. Jones GM, Trounson AO, Gardner DK, Kausche A, Lolatgis N, Wood C. Evolution of a culture protocol for successful blastocyst development and pregnancy. Hum Reprod 1998;13:169-77. https://doi.org/10.1093/humrep/13.1.169
  3. Menezo Y, Nicollet B, Herbaut N, Andre D. Freezing cocultured human blastocysts. Fertil Steril 1992;58:977-80. https://doi.org/10.1016/S0015-0282(16)55445-4
  4. Bonsi A. Blastocyst transfer and freezing: can this help us to improve the success of assisted reproduction? Singapore J Obstet Gynecol 1995;26:13-7.
  5. Kaufman RA, Menezo Y, Hazout A, Nicollet B, DuMont M, Servy EJ. Cocultured blastocyst cryopreservation: experience of more than 500 transfer cycles. Fertil Steril 1995;64:1125-9. https://doi.org/10.1016/S0015-0282(16)57972-2
  6. Nakayama T GY, Kanzaki H, Takabatake K, Himeno T, Takakura K, et al. Cryopreservation of human blastocyst. In: IX world congress on in vitro Fertilization and Alternated Assisted Reproduction, 1995 April 3-7; Vienna, Austria. p. 451-4.
  7. Cohen J, Simons RF, Edwards RG, Fehilly CB, Fishel SB. Pregnancies following the frozen storage of expanding human blastocysts. J In vitro Fert Embryo Transf 1985;2:59-64. https://doi.org/10.1007/BF01139337
  8. Saito N, Imai K, Tomizawa M. Effect of sugars-addition on the survival of vitrified bovine blastocysts produced in vitro. Theriogenology 1994;41:1053-60. https://doi.org/10.1016/S0093-691X(05)80028-7
  9. Yang NS, Lu KH, Gordon I, Polge C. Vitrification of blastocysts produced in vitro. Theriogenology 1992;37:326. https://doi.org/10.1016/0093-691X(92)90395-8
  10. Fahy GM, MacFarlane DR, Angell CA, Meryman HT. Vitrification as an approach to cryopreservation. Cryobiology 1984;21:407-26. https://doi.org/10.1016/0011-2240(84)90079-8
  11. 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;98:139-45. https://doi.org/10.1530/jrf.0.0980139
  12. Vanderzwalmen P, Delval A, Chatziparasidou A, Bertin G, Ectors F, Lejeune B, et al. Pregnancies after vitrification of human day 5 embryos. Hum Reprod 1997; 12(Suppl): 98.
  13. Van Der Zwalmen P, Gaurois B, Ectors FJ, Touati K, Massip A, Ectors F. Some factors affecting successful vitrification of mouse blastocysts. Theriogenology 1988;30:1177-83. https://doi.org/10.1016/0093-691X(88)90293-2
  14. Menezes DM, Orth D. Comparative analysis of changes resulting from bite plate therapy and Begg treatment. Angle Orthod 1975;45:259-66.
  15. Rall WF, Fahy GM. Ice-free cryopreservation of mouse embryos at -196 degrees C by vitrification. Nature 1985;313:573-5. https://doi.org/10.1038/313573a0
  16. Scheffen B, Van Der Zwalmen P, Massip A. A simple and efficient procedure for preservation of mouse embryos by vitrification. Cryo Letters 1986;7:260-9.
  17. Miyake T, Kasai M, Zhu SE, Sakurai T, Machida T. Vitrification of mouse oocytes and embryos at various stages of development in an ethylene glycol-based solution by a simple method. Theriogenology 1993;40:121-34. https://doi.org/10.1016/0093-691X(93)90346-7
  18. Carroll J, Depypere H, Matthews CD. Freeze-thaw-induced changes of the zona pellucida explains decreased rates of fertilization in frozen-thawed mouse oocytes. J Reprod Fertil 1990;90:547-53. https://doi.org/10.1530/jrf.0.0900547
  19. Germond M, Senn A, Rink K, Delacretas G, De Grandi P. Is assisted hatching of frozen-thawed embryos enhancing pregnancy outcome in patients who had several previous indation failures? J Reprod Fertil 1995;3:41-2.
  20. Vanderzwalmen P, Bertin G, Debauche C, Standaert V, van Roosendaal E, Vandervorst M, et al. Births after vitrification at morula and blastocyst stages: effect of artificial reduction of the blastocoelic cavity before vitrification. Hum Reprod 2002;17:744-51. https://doi.org/10.1093/humrep/17.3.744
  21. Son WY, Yoon SH, Yoon HJ, Lee SM, Lim JH. Pregnancy outcome following transfer of human blastocysts vitrified on electron microscopy grids after induced collapse of the blastocoele. Hum Reprod 2003;18:137-9. https://doi.org/10.1093/humrep/deg029
  22. Hiraoka K, Hiraoka K, Kinutani M, Kinutani K. Blastocoele collapse by micropipetting prior to vitrification gives excellent survival and pregnancy outcomes for human day 5 and 6 expanded blastocysts. Hum Reprod 2004;19:2884-8. https://doi.org/10.1093/humrep/deh504
  23. Mukaida T, Oka C, Goto T, Takahashi K. Artificial shrinkage of blastocoeles using either a micro-needle or a laser pulse prior to the cooling steps of vitrification improves survival rate and pregnancy outcome of vitrified human blastocysts. Hum Reprod 2006;21:3246-52. https://doi.org/10.1093/humrep/del285
  24. Martino A, Songsasen N, Leibo SP. Development into blastocysts of bovine oocytes cryopreserved by ultra-rapid cooling. Biol Reprod 1996;54:1059-69. https://doi.org/10.1095/biolreprod54.5.1059
  25. Lane M, Bavister BD, Lyons EA, Forest KT. Containerless vitrification of mammalian oocytes and embryos. Nat Biotechnol 1999;17:1234-6. https://doi.org/10.1038/70795
  26. Vanderzwalmen P, Bertin G, Debauche C, 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;18:1504-11. https://doi.org/10.1093/humrep/deg298
  27. 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. https://doi.org/10.1016/S1472-6483(10)61169-8
  28. Jo DH, Ko GR, Jung JH, Choi JR, Joo JK, Lee KS. Effects of the artificial shrinkage and assisted hatching before vitrification on the development of the vitrified mouse expanding blastocysts. Korean J Reprod Med 2008;35:275-83.
  29. Willadsen SM, Polge C, Rowson LE, Moor RM. Deep freezing of sheep embryos. J Reprod Fertil 1976;46:151-4. https://doi.org/10.1530/jrf.0.0460151
  30. Whittingham DG, Wood M, Farrant J, Lee H, Halsey JA. Survival of frozen mouse embryos after rapid thawing from -196 degrees C. J Reprod Fertil 1979;56:11-21. https://doi.org/10.1530/jrf.0.0560011
  31. Rall WF, Reid DS, Farrant J. Innocuous biological freezing during warming. Nature 1980;286:511-4. https://doi.org/10.1038/286511a0
  32. Smorag Z, Katska L, Wierzchos E. Some factors affecting the viability of mouse and cattle embryos frozen to -40 deg C before transfer to liquid nitrogen. Anim Reprod Sci 1981;4:65-72. https://doi.org/10.1016/0378-4320(81)90021-X
  33. Massip A, Vanderzwalmen P. In vitro survival of mouse embryos frozen in glycerol or glycerol-sucrose. Cryo Letters 1982;3:326.
  34. Szell A, Shelton JN. Osmotic and cryoprotective effects of glycerol- sucrose solutions on day-3 mouse embryos. J Reprod Fertil 1987;80:309-16. https://doi.org/10.1530/jrf.0.0800309
  35. Szell A, Shelton JN. Sucrose dilution of glycerol from mouse embryos frozen rapidly in liquid nitrogen vapour. J Reprod Fertil 1986;76:401-8. https://doi.org/10.1530/jrf.0.0760401
  36. Willadsen S, Polge C, Rowson LE. The viability of deep-frozen cow embryos. J Reprod Fertil 1978;52:391-3. https://doi.org/10.1530/jrf.0.0520391
  37. Friedler S, Giudice LC, Lamb EJ. Cryopreservation of embryos and ova. Fertil Steril 1988;49:743-64. https://doi.org/10.1016/S0015-0282(16)59879-3
  38. Leibo SP. Principles of cryopreservation for ART laboratory. In: Proceeding for 13th annual postgraduate program for ASRM. Course VII. Current techniques and new prontiers in cryopreservation and micromanipulation, 1997. p. 1-14.
  39. Sun ST, Choi JR, Son JB, Joo JK, Ko GR, Lee KS. The effect of long zona dissection using ICSI pipettes for mechanical assisted hatching in vitrified-thawed blastocyst transfers. J Assist Reprod Genet 2012;29:1431-4. https://doi.org/10.1007/s10815-012-9872-1

Cited by

  1. Morphoanatomical aspects of the starting material for the improvement of pineapple cryopreservation by the droplet-vitrification technique vol.93, pp.1, 2014, https://doi.org/10.1590/0001-3765202120190555