Post-thaw development of in vitro produced buffalo embryos cryopreserved by cytoskeletal stabilization and vitrification.
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 CO(2) incubator for 40 min for microfilament stabilization and were cryopreserved by a two-step vitrification method at 24 degrees 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.
Manjunatha
B M
BM
Department of Animal Reproduction, Veterinary College, Karnataka Veterinary, Animal and Fishery Sciences University, Bangalore, India. drmanjunathvet@gmail.com
Ravindra
J P
JP
Gupta
P S
PS
Devaraj
M
M
Honnappa
T G
TG
Krishnaswamy
A
A
D008841:Actin Cytoskeleton / Q000187:drug effects* / Q000502:physiology; D000818:Animals; D002020:Buffaloes / Q000196:embryology*; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D046149:Embryo Culture Techniques / Q000379:methods / Q000662:veterinary*; D019855:Ethylene Glycol / Q000494:pharmacology; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D008297:Male; D011247:Pregnancy
D003451:Cryoprotective Agents; D003571:Cytochalasin B; D019855:Ethylene Glycol; D004121:Dimethyl Sulfoxide