Effect of vitrification and slow freezing on in-vitro matured prepubertal goat oocytes

RAJNI SINHA; S D KHARCHE; N K SINHA; A K GOEL; S K JINDAL; SONIA SARASWAT

doi:10.56093/ijans.v84i3.38711

Authors

RAJNI SINHA Central Institute for Research on Goats, Makhdoom, Uttar Pradesh 281 122 India
S D KHARCHE Central Institute for Research on Goats, Makhdoom, Uttar Pradesh 281 122 India
N K SINHA Central Institute for Research on Goats, Makhdoom, Uttar Pradesh 281 122 India
A K GOEL Central Institute for Research on Goats, Makhdoom, Uttar Pradesh 281 122 India
S K JINDAL Central Institute for Research on Goats, Makhdoom, Uttar Pradesh 281 122 India
SONIA SARASWAT Central Institute for Research on Goats, Makhdoom, Uttar Pradesh 281 122 India

https://doi.org/10.56093/ijans.v84i3.38711

Keywords:

Cleavage, Cryopreservation, Oocytes, Prepubertal goats, Vitrification

Abstract

The present study was conducted to study the post thaw recovery rate of morphologically normal matured oocytes subjected to vitrification and slow freezing method. Oocytes were matured in TCM-199, which comprises sodium pyruvate (0.25 mM), L- glutamine (1 mg/10 ml), gentamycin (50 μg/ml) and supplemented with 10% fetal bovine serum at 38.5°C in CO₂ incubator under humidified air. After 27 h of in-vitro maturation oocytes were assigned for freezing, viz. vitrification, slow cooling and control cooling. The percentage of oocytes recovered after thawing was significantly higher for control than those for vitrification (69.67%) and slow freezing (85.00%), respectively. Moreover, the cleavage rate in vitrification, slow freezing and control were 1.14, 3.92 and 13.37%, respectively. Thus, the cleavage rate of oocytes in vitrification and slow freezing method did not differ significantly. However, there was a significant difference in cleavage rates of freezing groups and the control groups.

Downloads

Download data is not yet available.

References

Abe Y, Hara K, Matsumoto H, Kobayashi J, Sasada H, Ekwall H, Rodriguez-Martinez H and Sato E. 2005. Feasibility of a nylon- mesh holder for vitrification of bovine germinal vesicle oocytes in subsequent production of viable blastocysts. Biology of Reproduction 72: 1416–20.

Al Yacoub A N, Gauly M and Holtz W. 2010. Open pulled straw vitrification of goat embryos at various stages of development. Theriogenology 73: 1018–23.

Begin I, Bhatia B, Baldassarre H, Dinnyes A and Keefer L C. 2003. Cryopreservation of goat oocytes and in vitro derived 2-to 4 cell embryos using the cryoloop (CLV) and solid surface vitrification (SSLV) methods. Theriogenology 59: 1839–50.

Begin I, Bhatia B, Baldassarre H, Dinnyes A, Keefer C L.2003. Cryopreservation of goat oocytes and in vivo derived 2- to 4– cell embryos using the cryoloop (CLV) and solid-surface vitrification (SSV) methods. Theriogenology 59: 1839–50.

Berlinguer F, Succu S, Mossa F, Madeddu M, Bebbere D, Leoni G G and Naitana S. 2007. Effects of trehalose co-incubation on in vitro matured prepubertal ovine oocyte vitrification. Cryobiology 55: 27–34.

Fasano G, Vannin AS, Biramane J, Delbaere A and Englert Y. 2010. Cryopreservation of human failed maturation oocytes shows that vitrification gives superior outcomes to slow cooling. Cryobiology 61: 243–47.

Gasparrini B, Attanasio L, Anna R D, Monaco E and Rosella P D. 2007. Cryopreservation of in vitro matured buffalo oocytes by minimum volumes vitrification methods. Animal Reproduction Science 98: 335–42.

Hou-Yun P, Yun-Ping D, Shin-EN Z, Hua-Bin Z, Tong-Yi Wu, Guo-Chun G, Hai-ping W, Li-Li W, Ying L, Rong li, Rong W and Ning Li. 2005. Bovine oocytes vitrified by the open pulled straw method and used for somatic cell cloning supported development to term. Theriogenology 64: 1381–91.

Kharche S D, Goel A K, Jindal S K, Sinha N K and Yadav P. 2008a. Effect of somatic cells co-culture on cleavage and development of in vitro fertilized embryos. Indian Journal of Animal Sciences 78: 686–92.

Kharche S D, Yadav EN, Goel A K, Jindal S K and Sinha N K. 2008b. Influence of culture media on in vitro fertilization of goat oocytes. Indian Journal of Animal Sciences 78: 1075– 77.

Kharche S D, Goel A K, Jindal S K,Goel P and Jha B P. 2011. Birth of twin kids following transfer of in-vitro produced goat embryos. Indian Journal of Animal Sciences 81: 1132– 34.

Larman M G, Sheehanm C B and Gardner D K. 2006. Calcium- free vitrification reduces cryoprotectant-induced zona pellucida hardening and increases fertilization rates in mouse oocytes. Reproduction 131: 53–61.

Lins R D Pereira C S and Hünenberger P H. 2004. Trehalose protein interaction in aqueous solution. Proteins 55: 177–186. Martino A, Mogas M T, Palamo M J and Paramino M T. 1994. Meiotic competence of prepubertal goat oocytes. Theriogenology 41: 969.

Men H S, Chen J C, Shang E Y, Yang SC and Zou R J. 1996. Cryopreservation of Kumming mouse oocytes using slow cooling ultrarapid cooling and vitrification protocols. Theriogenology 47: 1423–31.

Otoi S, Kondo T S and Suzuki T. 1992. Developmental capacity of bovine oocytes cryopreseerved after maturation in vitro and of frozen thawed bovine embryos derived from frozen mature oocytes. Theriogenology 38: 711–19.

Otoi T, Yammoto K, Koyama N, Tachikawa S and Suzuki T. 1998. Cryopreservation of mature bovine oocytes by vitrification in straws. Cryobiology 37: 77–85.

Purohit G N, Meena H and Solanki K. 2012. Effect of vitrification on immature and in vitro matured, denuded and cumulus compact goat oocytes and their subsequent fertilization. Journal of Reproduction and Infertility 13: 53–59.

Rama Raju G A, Murali Krishna K, Prakash, G J and Madan K. 2006. Vitrification: An emerging technique for cryopreservation in assisted reproduction programmes. Embryo Talk Vol 1.4.

Saunders K M and Parks J E. 1999. Effects of cryopreservation procedures on the cytology and fertilization rate of in vitro- matured bovine oocytes. Biology of Reproduction 61: 178– 87.

Sharma G T, Kharche S D and Majumdar AC. 2006. Vitrification of in vitro matured goat oocytes and the effect on in vitro fertilization. Small Ruminant Research 64: 82–86.

Snedecor G W and Cochran W G. 1989. Statistical Methods. 6thedn. Iowa State University Press, USA.

Succu S, Leoni G G, Bebbere D, Berlinguer F, Mossa F, Bogliolo L, Madeddu M, Ledda S and Naitana S. 2007a. Vitrification devices affect structural and molecular status of in vitro matured ovine oocytes. Molecular Reproduction and Development 74: 1337–44.

Succu S, Leoni G S, Berlinguer F, Madeddu M, Bebbere D, Mossa F, Bogliolo L, Ledda S and Naitana S. 2007b. Effect of vitrification solutions and cooling upon in vitro matured prepubertal ovine oocytes. Theriogenology 68: 107–14.