Register      Login
Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

Effect of type of cryoprotectant on morphology and developmental competence of in vitro-matured buffalo (Bubalus bubalis) oocytes subjected to slow freezing or vitrification

S. K. Gautam A , V. Verma A , P. Palta A , M. S. Chauhan A and R. S. Manik A B
+ Author Affiliations
- Author Affiliations

A Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132 001, India.

B Corresponding author. Email: manik_rs@rediffmail.com

Reproduction, Fertility and Development 20(4) 490-496 https://doi.org/10.1071/RD07203
Submitted: 7 November 2007  Accepted: 4 February 2008   Published: 11 April 2008

Abstract

The present study examined the effects of different cryoprotectants on morphology and developmental competence of in vitro-matured buffalo oocytes after slow freezing or vitrification. After slow freezing in dimethyl sulfoxide (DMSO), ethylene glycol (EG) or 1,2-propanediol (PROH), at 1.0 or 1.5 m each, the proportion of morphologically normal oocytes recovered was significantly higher (P < 0.05) with 1.5 than 1.0 m for all cryoprotectants and was highest (P < 0.05) for 1.5 m DMSO. Following vitrification, the percentage of morphologically normal oocytes recovered was lower (P < 0.01) for 40% EG than for 40% DMSO, 20% EG + 20% DMSO or 20% EG + 20% PROH. The most common damage, irrespective of the cryopreservation method, was loss of cumulus mass. The cleavage rate and the proportion of vitrified–warmed oocytes that developed to morulae/blastocysts were significantly higher (P < 0.01) for 20% EG + 20% DMSO than for the other groups. A higher proportion of oocytes developed to morulae (11.5% v. 4.3%) or blastocysts (5.4% v. 0.6%) after vitrification in 20% EG + 20% DMSO than after slow freezing in 1.5 m DMSO. In conclusion, vitrification was more effective than slow freezing for the cryopreservation of in vitro-matured buffalo oocytes.


References

Aigner, S. , Van der Elst, J. , Siebzehnrubl, E. , Wildt, L. , Lang, N. , and Van Steirteghem, A. C. (1992). The influence of slow and ultra-rapid freezing on the organization of the meiotic spindle of the mouse oocytes. Hum. Reprod. 7, 857–864.
PubMed |

Almeida, P. A. , and Bolton, V. N. (1995). The effect of temperature fluctuations on the cytoskeletal organization and chromosomal constitution of the human oocyte. Zygote 3, 357–365.
PubMed |

Albarracin, J. L. , Morato, R. , Rojas, C. , and Mogas, T. (2005). Effect of vitrification in open pulled straws on the cytology of in vitro matured prepubertal and adult bovine oocytes. Theriogenology 63, 890–901.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Aman, R. R. , and Parks, J. E. (1994). Effects of cooling and rewarming on the meiotic spindle and chromosomes of in vitro matured bovine oocytes. Biol. Reprod. 50, 103–110.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Asada, M. , and Fukui, Y. (2000). Effect on fertilization and development by re-culture after freezing and thawing of bovine oocytes matured in vitro. Theriogenology 54, 889–898.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Chauhan, M. S. , Singla, S. K. , Palta, P. , Manik, R. S. , and Madan, M. L. (1998). In vitro maturation and fertilization, and subsequent development of buffalo (Bubalus bubalis) embryos: effects of oocyte quality and type of serum. Reprod. Fertil. Dev. 10, 173–177.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Dhali, A. , Manik, R. S. , Singla, S. K. , and Palta, P. (2000). Post-vitrification survival and in vitro maturation rate of buffalo (Bubalus bubalis) oocytes: effect of ethylene glycol concentration and exposure time. Anim. Reprod. Sci. 63, 159–165.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Fuku, E. , Kozima, T. , Shioya, Y. , Mercus, G. J. , and Downey, B. R. (1992). In vitro fertilization and development of frozen–thawed bovine oocytes. Cryobiology 29, 485–492.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Fuku, E. , Xia, L. , and Downey, B. R. (1995). Ultrastructural changes in bovine oocytes cryopreserved by vitrification. Cryobiology 32, 139–156.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Gasparrini, B. (2002). In vitro embryo production in buffalo species: state-of-the-art. Theriogenology 57, 237–256.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Gasparrini, B. , Attanasio, L. , De Rosa, A. , Monaco, E. , Di Palo, R. , and Campanile, G. (2007). Cryopreservation of in vitro matured buffalo (Bubalus bubalis) oocytes by minimum volumes vitrification methods. Anim. Reprod. Sci. 98, 335–342.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Gautam, S. K. , Verma, V. , Singh, B. , Palta, P. , Singla, S. K. , Chauhan, M. S. , and Manik, R. S. (2008). Effect of slow freezing on morphology and developmental competence of buffalo (Bubalus bubalis) immature oocytes. Anim. Reprod. Sci. 105, 311–318.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Gupta, V. , Manik, R. S. , Chauhan, M. S. , Singla, S. K. , Akshey, Y. S. , and Palta, P. (2006). Repeated ultrasound-guided transvaginal oocyte retrieval from cyclic Murrah buffaloes (Bubalus bubalis): oocyte recovery and quality. Anim. Reprod. Sci. 91, 89–96.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Im, K. S. , Kang, J. K. , and Kim, H. S. (1997). Effects of cumulus cells, different cryoprotectants, various maturation stages and preincubation before insemination on developmental capacity of frozen–thawed bovine oocytes. Theriogenology 47, 881–891.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Kumar, D. , Palta, P. , Manik, R. S. , Singla, S. K. , and Chauhan, M. S. (2007). Effect of culture media and serum supplementation on the development of in vitro fertilized buffalo embryos. Indian J. Anim. Sci. 77, 697–701.


Le Gal, F. , De Roover, R. , Verhaeghe, B. , Etienne, D. , and Massip, A. (2000). Development of vitrified matured cattle oocytes after thawing and culture in vitro. Vet. Rec. 146, 469–471.
PubMed |

Lim, J. M. , Ko, J. J. , Hwang, H. M. , and Niwa, K. (1999). Development of in vitro matured bovine oocytes after cryopreservation with different cryoprotectants. Theriogenology 51, 1303–1310.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Martino, A. , Songasen, N. , and Leibo, S. P. (1996). Development into blastocysts of bovine oocytes cryopreserved by ultra-rapid cooling. Biol. Reprod. 54, 1059–1069.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Mavrides, A. , and Morroll, D. (2002). Cryopreservation of bovine oocytes: is cryoloop vitrification the future to preserving the female gamete? Reprod. Nutr. Dev. 42, 73–80.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Otoi, T. , Tachikawa, S. , Kondo, S. , and Suzuki, T. (1993). Developmental capacity of bovine oocytes frozen in different cryoprotectants. Theriogenology 40, 801–807.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Otoi, T. , Tachikawa, S. , Kondo, S. , and Suzuki, T. (1994). Developmental competence of bovine oocytes frozen at different cooling rates. Cryobiology 31, 344–348.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Otoi, T. , Yamamoto, K. , Koyama, N. , and Suzuki, T. (1995). In vitro fertilization and development of immature and mature bovine oocytes cryopreserved by ethylene glycol with sucrose. Cryobiology 32, 455–460.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Otoi, T. , Yamamoto, K. , Koyama, N. , Tachikawa, S. , and Suzuki, T. (1998). Cryopreservation of mature bovine oocytes by vitrification in straws. Cryobiology 37, 77–85.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Palta, P. , and Chauhan, M. S. (1998). Laboratory production of buffalo (Bubalus bubalis) embryos. Reprod. Fertil. Dev. 10, 379–391.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Saunders, K. M. , and Parks, J. E. (1999). Effects of cryopreservation procedures on the cytology and fertilization rate of in vitro matured bovine oocytes. Biol. Reprod. 61, 178–187.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Schellander, K. , Peli, S. , Schmoli, F. , and Brem, G. (1994). Effects of different cryoprotectants and carbohydrates on freezing of mature and immature bovine oocytes. Theriogenology 42, 909–915.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Vajta, G. , Holm, P. , Kuwayama, M. , Booth, P. J. , Jacobsen, H. , Greve, T. , and Callesen, H. (1998). Open pulled straw (OPS) vitrification: a new way to reduce cryoinjuries of bovine oocytes and embryos. Mol. Reprod. Dev. 51, 53–58.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Wani, N. A. , Misra, A. K. , and Maurya, S. N. (2004). Effect of cryoprotectants and their concentration on in vitro development of vitrified–warmed immature oocytes in buffalo (Bubalus bubalis). Theriogenology 61, 831–842.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Zenzes, M. T. , Bielecki, R. , Casper, R. F. , and Leibo, S. P. (2001). Effects of chilling to 0°C on the morphology of meiotic spindles in human metaphase II oocytes. Fertil. Steril. 75, 769–777.
Crossref | GoogleScholarGoogle Scholar | PubMed |