CSIRO Publishing blank image blank image blank image blank imageBooksblank image blank image blank image blank imageJournalsblank image blank image blank image blank imageAbout Usblank image blank image blank image blank imageShopping Cartblank image blank image blank image You are here: Journals > Reproduction, Fertility and Development   
Reproduction, Fertility and Development
  Vertebrate Reproductive Science & Technology
 
blank image Search
 
blank image blank image
blank image
 
  Advanced Search
   

Journal Home
About the Journal
Editorial Board
Contacts
Content
Online Early
Current Issue
Just Accepted
All Issues
Special Issues
Research Fronts
Sample Issue
For Authors
General Information
Instructions to Authors
Submit Article
Open Access
For Referees
Referee Guidelines
Review an Article
Annual Referee Index
For Subscribers
Subscription Prices
Customer Service
Print Publication Dates

blue arrow e-Alerts
blank image
Subscribe to our email Early Alert or RSS feeds for the latest journal papers.

red arrow Connect with us
blank image
facebook twitter youtube

red arrow Connect with SRB
blank image
facebook TwitterIcon

Affiliated Societies

RFD is the official journal of the International Embryo Transfer Society and the Society for Reproductive Biology.



Article << Previous     |     Next >>   Contents Vol 22(1)

120 SUCCESSFUL LAMBING AFTER SUPER-COOLING ULTRARAPID VITRIFICATION SHEEP MORULAE

Y. M. Toishibekov A and H. D. Blackburn B

A Institute of Experimental Biology, Almaty, Kazakhstan;
B National Center of Genetic Resource Preservation, Fort Collins, CO, USA
   

Abstract
Export Citation
Print
  


Abstract

The aim of this work was to establish alternative methods for sheep morulae cryopreservation by using vitrification by open pulled straw (OPS) methods and super-cooling ultra-rapid vitrification (SCURV). Both treatments used a vitrification solution (VS) of 20% (3.6 mol L-1) ethylene glycol (EG), 20% (2.4 mol L-1) dimethylsulfoxide (DMSO), 0.5 mol L-1 sucrose in DPBS with 10% BSA in both methods. In our experiment we used the Vit-Master™ (MTG, Germany). The super-cooled LN facilitates heat transmission between LN and the cryosolution interface, and this is efficient for bovine semen and blastocyst cryoconservation (Arav et al. 2002). By surgical flushing of 24 super stimulated ewes 121 transferrable morulae were harvested; 30 morulae were transferred fresh to synchronised recipients and the others were cryopreserved by OPS (n = 49) or SCURV (n = 42), respectively thawed or warmed, and transferred to recipients. Embryos were vitrified using the OPS method. They were first incubated in 50% VS for 2 min and then transferred for 30 s into 100% VS. Each embryo was loaded by touching a 1-μL drop with the straw, which was immediately submerged into and stored in liquid nitrogen. Warming was done by placing the narrow end of the straw into DPBS + 0.25M sucrose for 5 min. Embryos were then transferred into DPBS + 0.125 M sucrose for 3 min and finally to DPBS until transfer. The SCURV morulae were then exposed to 50 and 100% VS at 37°C for 2 min and 30 s, respectively. Embryos after saturation VS have been transferred by on a surface of a nylon loop (volume 20 μL, diameter 0.5 mm) and using negative pressure temperature of liquid nitrogen in the chamber for freezing with the VIT-Master. Thawing vitrified embryos was accomplished by placing the vitrified embryos in solutions of sucrose 0.25 and 0.125 with expositions of 2 and 3 min, accordingly. After embryos were thawed, only good quality embryos were transferred. Importantly, our data suggest that by using the SCURV method, the toxic elements contained in the cryopreservation solution can be reduced while maintaining a similar ability to produce viable morulae for implantation as the OPS method. Although further work on the developmental competence of embryos cryopreserved with the SCURV method are needed, these data suggest that the faster freeze rate and lower levels of cryoprotectants of SCURV are able to minimize ice crystal formation and should be further evaluated as a routine mechanism for cryopreserving sheep morulae.

Reproduction, Fertility and Development 22(5305) 218–219   http://dx.doi.org/10.1071/RDv22n1Ab120
Published online: 08 December 2009




 
Top  Email this page
 
   


Legal & Privacy | Contact Us | Help

CSIRO

© CSIRO 1996-2014