期刊论文详细信息
Journal of Animal Science and Biotechnology
An efficient method for the sanitary vitrification of bovine oocytes in straws
Shien Zhu1  Yunpeng Hou3  Yi Fang1  Baoyu Jia1  Guangbin Zhou2  Xiangwei Fu1  Yanhua Zhou1 
[1] National Engineering Laboratory for Animal Breeding, Key Laboratory of Animal genetics, Breeding and Reproduction, Ministry of Agriculture, College of Animal Science and Technology, China Agricultural University, Beijing 100193, P.R. China;Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University (Chengdu Campus), Wenjiang 611130, P.R. China;State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, P.R. China
关键词: Vitrification;    Straw;    Oocytes;    Cryopreservation;    Bovine;   
Others  :  803498
DOI  :  10.1186/2049-1891-5-19
 received in 2014-02-18, accepted in 2014-04-08,  发布年份 2014
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【 摘 要 】

Background

At present, vitrification has been widely applied to humans, mice and farm animals. To improve the efficiency of vitrification in straw, bovine oocytes were used to test a new two-step vitrification method in this study.

Results

When in vitro matured oocytes were exposed to 20% ethylene glycol (EG20) for 5 min and 40% ethylene glycol (EG40) for 30 s, followed by treatment with 30% glycerol (Gly30), Gly40 or Gly50, a volume expansion was observed in Gly30 and Gly40 but not Gly50. This indicates that the intracellular osmotic pressure after a 30 s differs between EG40 and ranged between Gly40 (approximately 5.6 mol/L) and Gly50 (approximately 7.0 mol/L). Since oocytes are in EG40 just for only a short period of time (30 s) and at a lower temperature (4°C), we hypothesize that the main function of this step in to induce dehydration. Based on these results, we omitted the EG40 step, before oocytes were pretreated in EG20 for 5 min, exposed to pre-cooled (4°C) Gly50, for 30 s, and then dipped into liquid nitrogen. After warming, 81.1% of the oocytes survived, and the surviving oocytes developed into cleavage stage embryos (63.5%) or blastocysts (20.0%) after parthenogenetic activation.

Conclusions

These results demonstrate that in a two-step vitrification procedure, the permeability effect in the second step is not necessary. It is possible that the second step is only required to provide adequate osmotic pressure to condense the intracellular concentration of CPAs to a level required for successful vitrification.

【 授权许可】

   
2014 Zhou et al.; licensee BioMed Central Ltd.

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【 参考文献 】
  • [1]Rall WF, Fahy GM: Ice-free cryopreservation of mouse embryos at -196°C by vitrification. Nat Commun 1985, 313:573-575.
  • [2]Kuwayama M, Vajta G, Kato O, Leibo SP: Highly efficient vitrification method for cryopreservation of human oocytes. Reprod Biomed Online 2005, 11:300-308.
  • [3]Vanderzwalmen P, Ectors F, Grobet L, Prapas Y, Panagiotidis Y, Vanderzwalmen S, Stecher A, Frias P, Liebermann J, Zech NH: Aseptic vitrification of blastocysts from infertile patients, egg donors and after IVM. Reprod Biomed Online 2009, 19:700-707.
  • [4]Vanderzwalmen P, Connan D, Grobet L, Wirleitner B, Remy B, Vanderzwalmen S, Zech N, Ectors FJ: Lower intracellular concentration of cryoprotectants after vitrification than after slow freezing despite exposure to higher concentration of cryoprotectant solutions. Hum Reprod 2013, 28:2101-2110.
  • [5]Liang W, Jun L, Guangbin Z, Yunpeng H, Junjie L, Shien Z: Quantitative investigations on the effects of exposure durations to the combined cryoprotective agents on mouse oocyte vitrification procedures. Biol Reprod 2011, 85:884-894.
  • [6]Edashige K, Ohta S, Tanaka M, Kuwano T, Valdez DM, Hara T, Jin B, Takahashi S, Seki S, Koshimoto C: The role of aquaporin 3 in the movement of water and cryoprotectants in mouse morulae. Biol Reprod 2007, 77:365-375.
  • [7]Campos-Chillon LF, Walker DJ, De La Torre-Sanchez JF, Seidel GE Jr: In vitro assessment of a direct transfer vitrification procedure for bovine embryos. Theriogenology 2006, 65:1200-1214.
  • [8]Tachikawa S, Otoi T, Kondo S, Machida T, Kasai M: Successful vitrification of bovine blastocysts, derived by in vitro maturation and fertilization. Mol Reprod Dev 1993, 34:266-271.
  • [9]Vajta G, Holm P, Greve T, Callesen H: Factors affecting survival rates of in vitro produced bovine embryos after vitrification and direct in-straw rehydration. Anim Reprod Sci 1996, 45:191-200.
  • [10]Vajta G, Holm P, Kuwayama M, Booth PJ, Jacobsen H, Greve T, Callesen H: Open pulled straw (OPS) vitrification: a new way to reduce cryoinjuries of bovine ova and embryos. Mol Reprod Dev 1998, 51:53-58.
  • [11]Le Gal F, Massip A: Cryopreservation of cattle oocytes: effects of meiotic stage, cycloheximide treatment, and vitrification procedure. Cryobiology 1999, 38:290-300.
  • [12]Dinnyés A, Dai Y, Jiang S, Yang X: High developmental rates of vitrified bovine oocytes following parthenogenetic activation, in vitro fertilization, and somatic cell nuclear transfer. Biol Reprod 2000, 63:513-518.
  • [13]KATO O, NAGAI T: High survival rate of bovine oocytes matured in vitro following vitrification. J Reprod Dev 2004, 50:685-696.
  • [14]Pedro PB, Yokoyama E, Zhu SE, Yoshida N, Valdez DM Jr, Tanaka M, Edashige K, Kasai M: Permeability of mouse oocytes and embryos at various developmental stages to five cryoprotectants. J Reprod Dev 2005, 512:235-246.
  • [15]Wang X, Al Naib A, Sun D, Lonergan P: Membrane permeability characteristics of bovine oocytes and development of a step-wise cryoprotectant adding and diluting protocol. Cryobiology 2010, 61:58-65.
  • [16]Whittingham DG: Fertilization in vitro and development to term of unfertilized mouse oocytes previously stored at—196 C. J Reprod Fertil 1977, 49:89-94.
  • [17]Fuku E, Kojima T, Shioya Y, Marcus GJ, Downey BR: In vitro fertilization and development of frozen-thawed bovine oocytes. Cryobiology 1992, 29:485-492.
  • [18]Wani NA, Misra AK, Maurya SN: Maturation rates of vitrified-thawed immature buffalo (Bubalus bubalis) oocytes: effect of different types of cryoprotectants. Anim Reprod Sci 2004, 84:327-335.
  • [19]Yamada C, Caetano HVA, Simões R, Nicacio AC, Feitosa WB, Assumpção MEOD, Visintin JA: Immature bovine oocyte cryopreservation: comparison of different associations with ethylene glycol, glycerol and dimethylsulfoxide. Anim Reprod Sci 2007, 99:384-388.
  • [20]Sharma GT, Dubey PK, Chandra V: Morphological changes, DNA damage and developmental competence of in vitro matured, vitrified-thawed buffalo (Bubalus bubalis) oocytes: a comparative study of two cryoprotectants and two cryodevices. Cryobiology 2010, 60:315-321.
  • [21]Saha S, Otoi T, Takagi M, Boediono A, Sumantri C, Suzuki T: Normal calves obtained after direct transfer of vitrified bovine embryos using ethylene glycol, trehalose, and polyvinylpyrrolidone. Cryobiology 1996, 33:291-299.
  • [22]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, 98:139-145.
  • [23]Rosenkrans CF, Zeng GQ, McNamara GT, Schoff PK, First NL: Development of bovine embryos in vitro as affected by energy substrates. Biol Reprod 1993, 49:459-462.
  • [24]Hou Y, Dai Y, Zhu S, Zhu H, Wu T, Gong G, Wang H, Wang L, Liu Y, Li R: Bovine oocytes vitrified by the open pulled straw method and used for somatic cell cloning supported development to term. Theriogenology 2005, 64:1381-1391.
  • [25]Fujihira T, Nagai H, Fukui Y: Relationship between equilibration times and the presence of cumulus cells, and effect of taxol treatment for vitrification of in vitro matured porcine oocytes. Cryobiology 2005, 51:339-343.
  • [26]Van Wagtendonk-de Leeuw AD, Den Daas J, Rall WF: Field trial to compare pregnancy rates of bovine embryo cryopreservation methods: vitrification and one-step dilution versus slow freezing and three-step dilution. Theriogenology 1997, 48:1071-1084.
  • [27]Leibo SP, Mazur P, Jackowski SC: Factors affecting survival of mouse embryos during freezing and thawing. Exp Cell Res 1974, 89:79-88.
  • [28]Jin B, Mochida K, Ogura A, Koshimoto C, Matsukawa K, Kasai M, Edashige K: Equilibrium vitrification of mouse embryos at various developmental stages. Mol Reprod Dev 2012, 79:785-794.
  • [29]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, 89:91-97.
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