J Vet Sci.  2009 Jun;10(2):153-156. 10.4142/jvs.2009.10.2.153.

Post-thaw development of in vitro produced buffalo embryos cryopreserved by cytoskeletal stabilization and vitrification

• Manjunatha BM
• Ravindra J
• Gupta PS
• Devaraj M
• Honnappa T
• Krishnaswamy A

• KMID: 1102972
• DOI: http://doi.org/10.4142/jvs.2009.10.2.153

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 24degrees 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 min. After the initial exposure, the embryos were transferred to a 7 microl 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.

Keyword

buffalo; cytochalasin-b; embryo; vitrificationm

MeSH Terms

Animals
Buffaloes/*embryology
Cryopreservation/methods/*veterinary
Cryoprotective Agents/pharmacology
Cytochalasin B/pharmacology
Dimethyl Sulfoxide/pharmacology
Embryo Culture Techniques/methods/*veterinary
Ethylene Glycol/pharmacology
Female
Fertilization in Vitro/*veterinary
Male
Microfilaments/*drug effects/physiology
Pregnancy

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