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http://dx.doi.org/10.5653/cerm.2014.41.3.115

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)
Publication Information
Clinical and Experimental Reproductive Medicine / v.41, no.3, 2014 , pp. 115-119 More about this Journal
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
Artificial shrinkage; Blastocyst; Sucrose; Vitrification;
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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.   DOI   ScienceOn
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.   DOI   ScienceOn
3 Menezo Y, Nicollet B, Herbaut N, Andre D. Freezing cocultured human blastocysts. Fertil Steril 1992;58:977-80.   DOI
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.   DOI
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.   DOI   ScienceOn
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.   DOI   ScienceOn
9 Yang NS, Lu KH, Gordon I, Polge C. Vitrification of blastocysts produced in vitro. Theriogenology 1992;37:326.   DOI   ScienceOn
10 Fahy GM, MacFarlane DR, Angell CA, Meryman HT. Vitrification as an approach to cryopreservation. Cryobiology 1984;21:407-26.   DOI   ScienceOn
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.   DOI   ScienceOn
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.   DOI   ScienceOn
14 Menezes DM, Orth D. Comparative analysis of changes resulting from bite plate therapy and Begg treatment. Angle Orthod 1975;45:259-66.
15 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.   DOI   ScienceOn
16 Rall WF, Fahy GM. Ice-free cryopreservation of mouse embryos at -196 degrees C by vitrification. Nature 1985;313:573-5.   DOI   ScienceOn
17 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.
18 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.   DOI   ScienceOn
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.   DOI   ScienceOn
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.   DOI   ScienceOn
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.   DOI   ScienceOn
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.   DOI   ScienceOn
24 Martino A, Songsasen N, Leibo SP. Development into blastocysts of bovine oocytes cryopreserved by ultra-rapid cooling. Biol Reprod 1996;54:1059-69.   DOI   ScienceOn
25 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.
26 Lane M, Bavister BD, Lyons EA, Forest KT. Containerless vitrification of mammalian oocytes and embryos. Nat Biotechnol 1999;17:1234-6.   DOI   ScienceOn
27 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.   DOI   ScienceOn
28 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.   DOI   ScienceOn
29 Willadsen SM, Polge C, Rowson LE, Moor RM. Deep freezing of sheep embryos. J Reprod Fertil 1976;46:151-4.   DOI   ScienceOn
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.   DOI   ScienceOn
31 Rall WF, Reid DS, Farrant J. Innocuous biological freezing during warming. Nature 1980;286:511-4.   DOI   ScienceOn
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.   DOI   ScienceOn
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.   DOI   ScienceOn
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.   DOI   ScienceOn
36 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.   DOI
37 Willadsen S, Polge C, Rowson LE. The viability of deep-frozen cow embryos. J Reprod Fertil 1978;52:391-3.   DOI   ScienceOn
38 Friedler S, Giudice LC, Lamb EJ. Cryopreservation of embryos and ova. Fertil Steril 1988;49:743-64.   DOI
39 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.