Acta Vet. Brno 2012, 81: 229-234

http://dx.doi.org/10.2754/avb201281030229

Effect of cysteamine supplementation during in vitro culture of early stage bovine embryos on blastocyst rate and quality

Martina Lojkic1, Iva Getz1, Marko Samardzija1, Mario Matkovic2, Goran Bacic1, Tugomir Karadjole1, Nino Macesic1, Ivan Folnozic1, Branimira Spoljaric1

1University of Zagreb, Faculty of Veterinary Medicine, Clinic for Obstetrics and Reproduction, Zagreb, Croatia
2Center for Reproduction and Animals Breeding of Croatia, Novi Zagreb, Croatia

The aim of this study was to evaluate whether the addition of cysteamine to the in vitro culture media enhances the yield, hatching rate, total cell number and inner cell mass/total cell number ratio of bovine embryos. A total of 933 bovine oocytes collected from ovaries of 60 slaughtered donors were subjected to in vitro maturation and in vitro fertilization. Following fertilization, embryos were cultured in synthetic oviductal fluid without glucose. After 24 h embryos were transferred into synthetic oviductal fluid with 1.5 mM glucose and 0 (control), 50, 100 and 200 µM of cysteamine. After 48 h, the embryos were transferred into synthetic oviductal fluid with glucose but without cysteamine and cultured until Day 9. The number of cleaved embryos on Day 2, the total number of blastocysts on Day 7 and the number of hatched blastocysts on Day 9 were calculated. Differential staining of inner cell mass and trophectoderm cells of blastocysts were performed on Day 7 and Day 9 of in vitro culture. Supplementation of in vitro culture media with 100 µM cysteamine increased the blastocyst yield (P < 0.05) without affecting the hatching rate. Furthermore, the embryos cultured in the presence of 100 µM cysteamine had significantly higher number of inner cell mass cells (P < 0.05) and the proportion of inner cell mass cells (P < 0.05) compared with the controls. The results of the present study demonstrated that the addition of 100 µM cysteamine to the in vitro culture media improved blastocyst production rate and enhance embryo quality, which could lead to the improvement of the in vitro culture system for bovine embryos.

References

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