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Effects of Vitrification on Cumulus Expansion and Nuclear Maturation of Abattoir-Derived Cattle Oocytes
* 1, 2 , 2 , 1 , 2, 3 , 2 , 1, 4
1  Germplasm Conservation and Reproductive Biotechnologies, Agricultural Research Council, Animal Production, Irene, South Africa
2  Department of Animal, Wildlife and Grassland Sciences, University of the Free State, Bloemfontein, South Africa
3  Department of Agriculture, Directorate: Animal Production, Subdirectorate: Animal Improvement, Pretoria, South Africa
4  Department of Agriculture and Animal Health, College of Agriculture and Environmental Sciences, University of South Africa, Pretoria, South Africa
Academic Editor: Mohammed Gagaoua

Abstract:

The present study aimed to assess the impact of vitrification on cumulus cell growth and nuclear maturation in bovine oocytes at different meiotic stages. Oocytes were divided into four treatment groups: positive control (immature fresh oocytes), negative control (immature oocytes exposed to CPA), immature vitrified, and mature vitrified. Heterogeneous cattle ovaries of unknown reproductive status were collected from a local abattoir and transported (37oC) to the laboratory within 2 h of slaughter. The aspiration method was used to retrieve oocytes from the ovaries. Immature oocytes and mature oocytes were vitrified using a conventional straw vitrification method. The straws were pre-cooled by placing them horizontally on a Styrofoam rack above LN2 vapour. Cryopreserved oocytes were then subjected to warming or thawing media and then immature vitrified oocytes were incubated in maturation medium for 22 h. Oocytes were evaluated for cumulus cell growth and nuclear maturation. Treatment means were compared using a least significant difference test. A high percentage of oocytes with fully expanded cumulus cells was recorded in the control group (83.1±12.0) and mature vitrified (77.2±14.0) group compared to the immature vitrified group (43.8±15.0; p< 0.05). Immature oocytes exposed to CPA (16.7±9.6) and the immature vitrified group (22.8±10.3) recorded a high percentage of oocytes with no cumulus cells expansion following maturation (p< 0.05). A high percentage of oocytes reaching metaphase I was recorded for immature oocytes exposed to CPA (54.2±19.3) and mature vitrified oocytes (60.3±12.4; p< 0.05). Both the control group and immature vitrified group recorded a high percentage of oocytes reaching metaphase II (48.5±6.0; 42.3±7.8), respectively, compared to the matured vitrified oocytes (29.2±13.2; p< 0.05). In conclusion, the results demonstrate that vitrifying bovine oocytes during the immature stage results in a higher maturation rate post-warming, indicating that the immature stage is more suitable to effective oocyte cryopreservation.

Keywords: cattle oocytes; cumulus expansion, nuclear maturation; vitrification

 
 
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