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The Comparison of Survival Rates of Vitrified Mouse Blastocysts in Various Vitrification Solutions and Apparatuses  

Cho, Dong-Hyu (Department of Obstetrics and Gynecology, Chonbuk National University Medical School)
Lee, Ky-Sook (Department of Obstetrics and Gynecology, Chonbuk National University Medical School)
Rheu, Chul-Hee (Department of Obstetrics and Gynecology, Chonbuk National University Medical School)
Kwon, Jung-Kee (Laboratory Animal Medicine, College of Veterinary Medicine, Chonbuk National University)
Lee, Jeong-Heon (Department of Obstetrics and Gynecology, Chonbuk National University Medical School)
Publication Information
Journal of Veterinary Clinics / v.29, no.1, 2012 , pp. 27-32 More about this Journal
Abstract
The purpose of this study was to evaluate survival rates of vitrified mouse blastocysts in various vitrification solutions (cryoprotectants) and apparatuses. The mouse blastocysts were harvested from culture of mouse 2 cell embryo and were divided into three group (i) untreated (control); (ii) exposed to cryoprotectant agents; or (iii) cryopreserved by various vitrification apparatuses. Vitrification solutions are 40% ethylene glycol (EG) + 5.8 mg/mL ficoll + 0.5M sucrose (EFS solution), 3M glycerol + 3M EG (ES solution), 20% EG + 20% dimethyl sulfoxide (ED solution), 3M EG + 1.0 m sucrose (ES solution). Vitrification apparatuses consisted of 5 groups ; closed plastic straw (CPS), electron microscope (EM) grid, cryoloop, open pulled straw (OPS), and glass micropippete in plastic straw (GPS). The survival rates of control were 88.0%. The survival rates of exposed blastocysts in EFS, GE, ED, and ES solutions were 70.8%, 43.5% (P<0.01), 83.3% and 65.2%, respectively. The survival rates of vitrified blastocysts in CPS, EM grid, cryoloop, OPS and GPS were 56.5% (P< 0.01), 72.7%, 83.3%, 60.9% (P<0.05) and 54.2% (P<0.01), respectively. Among the vitrification solutions, the highest survival rate was seen in blastocysts vitrified in EG + DMSO (83.3%). The survival rate was not significantly different from that of the control (88%). Blastocysts cryopreserved with glycerol in all groups had an overall low survival rate of 43.5%. Survival rate of mouse blastocysts between vitrification apparatuses showed higher in cryoloop.
Keywords
cryoprotectants; blastocysts; vitrification;
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1 Tan X, Song E, Liu X, You W, Wan F. Factors affecting the survival, fertilization and embryonic development of mouse oocytes after vitrification using glass capillaries. In Vitro Cell Dev Bio Anim. 2009; 45: 420-429.   DOI   ScienceOn
2 Trounson A, Mohr L. Human Pregnancy following cryopreservation, thawing and transfer of an eight-cell embryo. Nature 1983; 305: 707-709.   DOI   ScienceOn
3 Wang L, Liu J, Zhou GB, Hou YP, Li JJ, Zhu SE. Quantitative investigations on the Effects of Exposure Durations to the Combined Cryoprotective Agents on Mouse Oocyte Vitrification Procedures. Bio Reprod 2011; Jun 22: 1-28.
4 Whittingham DG, Leibo SP, Mazur P. Survival of mouse embryos frozen to -196 degrees and -269 degrees C. Science 1972; 178: 411-414.   DOI
5 Yeoman RR, Gerami-Naini B, Mitalipov S, Nusser KD, Widmann-Browning AA, Wolf DP. Cryoloop vitrification yields superior survival of Rhesus monkey blastocysts. Hum Reprod 2001; 16: 1965-1969.   DOI
6 Cervera RP, Garcia-Ximénez F. Vitrification of zona-free rabbit expanded or hatching blastocysts: a possible model for human blastocysts. Hum Reprod 2003; 18: 2151-215.   DOI
7 Dumoulin JC, Bergers-Janssem JM, Pieters MH, Enqinsu ME, Geraedts JP, Evers JL. The protective effects of polymers in the cryopreservation of human and mouse zonae pellucidae and embryos. Feril Steril 1994; 62: 793-798.
8 Friedler S, Giudice LC, Lamb EJ. Cryopreservation of embryos and ova. Fertil Steril 1988; 49: 743-764.
9 Mahmoud KG, Scholkamy TH, Ahmed YF, Seidel GE Jr, Nawito MF. Effect of different combinations of cryoprotectants on in vitro maturation of immature buffalo (Bubalus bubalis) oocytes vitrified by straw and open-pulled straw methods. Reprod Domest Anim. 2010; 45: 565-571.
10 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: 121-134.   DOI   ScienceOn
11 Mullen GF, Li M, Li Y, Chen ZJ, Crister JK. Human oocyte vitrification the permiaility of metaphase II oocytes to water and ethylene glycol and appliance toward vitrification. Fertil Steril. 2008; 89: 1812-1825.   DOI   ScienceOn
12 Otoi T, Yamamoto K, Koyama N, Tachikawa S, Suzuki T. Cryopreservation of mature bovine oocytes by vitrification in straws. Cryobiology. 1998; 37: 77-85.   DOI   ScienceOn
13 Paynter SJ, Cooper A, Gregory L, Fuller BJ, Shaw RW. Permeability characteristics of human oocytes in the presence of the cryoprotectant dimethylsulphoxide. Hum Reprod 1999; 14: 2338-2342.   DOI
14 Paynter SJ, O'Neil L, Fuller BJ, Shaw RW. Membrane permeability of human oocytes in the presence of the cryoprotectant Propane-1,2-diol. Fertil Steril. 2001; 75: 532-538.   DOI   ScienceOn
15 Pegg DE. Principles of cryopreservation. Methods Mol Biol. 2007; 368: 39-57.
16 Rall WF, Fahy GM. Ice-free cryopreservation of mouse embryos at -196 degrees C by vitrification. Nature 1985; 313: 573-575.   DOI   ScienceOn
17 Rall W F, Wood M J, Kirby C, Whittingham D G. Development of mouse embryos cryopreserved by vitrification. J. Reprod. Fertil 1987; 80: 488-504.
18 Reubinoff BE, Pera MF, Vajta G, Trounson AO. Effective cryopreservation of human embryonic stem cells by the open pulled straw vitrification method. Hum Reprod 2001; 16:2187-2194.   DOI   ScienceOn
19 Schneider U, Mazur P. Relative influence of unfrozen fraction and salt concentration on the survival of slowly frozen eightcell mouse embryos. Cryobiology 1987; 24: 17-41.   DOI   ScienceOn
20 Shaw PW, Fuller BJ, Bernard A, Shaw RW. Vitrification of mouse oocytes: improved rates of survival fertilization and development to blastocysts. Mol Reprod Dev 1991; 29: 373- 378.   DOI   ScienceOn
21 Széll A, Shelton JN. Sucrose dilution of glycerol from mouse embryos frozen rapidly in liquid nitrogen vapour. J Reprod Fertil 1986; 76: 401-408.   DOI
22 Takahashi Y, Kanagawa H. Effect of equilibration period on the viability of frozen-thawed mouse morulae after rapid freezing. Mol Reprod Dev. 1990; 26: 105-110.   DOI   ScienceOn
23 Ali J, Shelton JN. Design of vitrification solutions for the cryopreservation of embryos. J Reprod Fertil. 1993; 99: 471-477.   DOI
24 Ashwood-Smith MJ, Morris GW, Fowler R, Appleton TC, Ashorn R. Physical factors are involved in the destruction of embryos and oocytes during freezing and thawing procedures. Hum Reprod 1988; 3: 795-802.
25 Aye M, Di Giorgio C, De Mo M, Botta A, Perrin J, Courbiere B. Assessment of the genotoxicity of three cryoprotectants used for human oocyte vitrification: dimethyl sulfoxide, ethylene glycol and propylene glycol. Food Chem Toxicol. 2010; 48: 1905-1912.   DOI   ScienceOn
26 Bernart W, Kamel M, Neulen J, Breckwoldt M. Influence of the developmental stage and the equilibration time on the outcome of ultrarapid cryopreservation of mouse embryos. Hum Reprod 1994; 9: 100-102.