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- Title
Post-thaw development of in vitro produced buffalo embryos cryopreserved by cytoskeletal stabilization and vitrification.
- Authors
Manjunatha, B. M.; Ravindra, J. P.; Gupta, P. S. P.; Devaraj, M.; Honnappa, T. G.; Krishnaswamy, A.
- 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 mm for microfilament stabilization and were cryopreserved by a two-step vitrification method at 24°C 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 mm. 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.
- Subjects
THAWING; ETHYLENE glycol; SUCROSE; DISACCHARIDES; ALKENES
- Publication
Journal of Veterinary Science, 2009, Vol 10, Issue 2, p153
- ISSN
1229-845X
- Publication type
Article
- DOI
10.4142/jvs.2009.10.2.153