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KMID : 0893420090100020153
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Journal of Veterinary Science
2009 Volume.10 No. 2 p.153 ~ p.156
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Post-thaw development of in vitro produced buffalo embryos cryopreserved by cytoskeletal stabilization and vitrification
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Manjunatha B. M.
Ravindra J. P.
Gupta P. S. P.
Devaraj M.
Honnappa T. G.
Krishnaswamy A.
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Abstract
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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 nonvitrified control. However, the hatching rate of cyto-btreated 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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KEYWORD
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buffalo, cytochalasin-b, embryo, vitrificationm
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