Post-thaw development of in vitro produced buffalo embryos cryopreserved by cytoskeletal stabilization and vitrification |
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| Authors: | B. M. Manjunatha J. P. Ravindra P. S. P. Gupta M. Devaraj T. G. Honnappa A. Krishnaswamy |
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| Affiliation: | 1Department of Animal Reproduction, Gynaecology and Obstetrics, Veterinary College, Karnataka Veterinary, Animal and Fishery Sciences University, Bangalore-560 024, India.;2National Institute of Animal Nutrition and Physiology, Indian Council of Agricultural Research, Adugodi, Bangalore-560 030, India. |
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| Abstract: | The present study was conducted to examine post-thaw in vitro developmental competence of buffalo embryos cryopreserved by cytoskeletal stabilization and vitrification. In vitro produced embryos were incubated with a medium containing cytochalasin-b (cyto-b) in a CO2 incubator for 40 min for microfilament stabilization and were cryopreserved by a two-step vitrification method at 24℃ in the presence of cyto-b. Initially, the embryos were exposed to 10% ethylene glycol (EG) and 10% dimethylsulfoxide (DMSO) in a base medium for 4 min. After the initial exposure, the embryos were transferred to a 7 µl drop of 25% EG and 25% DMSO in base medium and 0.3 M sucrose for 45 sec. After warming, the embryos were cultured in vitro for 72 h. The post-thaw in vitro developmental competence of the cyto-b-treated embryos did not differ significantly from those vitrified without cyto-b treatment. The hatching rates of morulae vitrified without cyto-b treatment was significantly lower than the non-vitrified control. However, the hatching rate of cyto-b-treated vitrified morulae did not differ significantly from the non-vitrified control. This study demonstrates that freezing of buffalo embryos by cytoskeletal stabilization and vitrification is a reliable method for long-term preservation. |
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| Keywords: | buffalo cytochalasin-b embryo vitrificationm |
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