Effect of reduced concentrations of glycerol and various macromolecules on the cryopreservation of mouse and cattle embryos.
The effect of different macromolecules [bovine serum albumin (BSA), Pluronic F-68, (ET surfactant), or sodium hyaluronate (SH)] on postthaw survival of mouse morulae and in vivo- and in vitro-derived bovine blastocysts frozen in 10, 5, or 1% glycerol solutions was investigated. Embryos were equilibrated with cryoprotectant solution at 25 degrees C for 10 min, seeded at -5 degrees C, cooled at 0.5 degrees C/min to -35 degrees C, and plunged into liquid nitrogen. Embryos were thawed in a 35 degrees C water bath, glycerol was removed with 0.6 M sucrose at 25 degrees C for 5 min, and postthaw viability was evaluated after 1, 24, and 48 h in culture. The addition of BSA supplementation improved postthaw survival of mouse morulae frozen in 5% glycerol, but not in 10% glycerol. All three macromolecular supplements were effective in increasing survival of mouse morulae in 5% glycerol but only BSA and SH were effective in increasing postthaw survival of in vivo- and in vitro-derived bovine blastocysts. None of the macromolecular supplements improved postthaw survival of embryos frozen in 1% glycerol.
Palasz
A T
AT
Department of Herd Medicine and Theriogenology, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5B4, Canada.
Thundathil
J
J
De La Fuente
J
J
Mapletoft
R J
RJ
D000818:Animals; D001755:Blastocyst / Q000187:drug effects*; D002417:Cattle / Q000196:embryology*; D002470:Cell Survival; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004624:Embryo Transfer; D005260:Female; D005307:Fertilization in Vitro; D005990:Glycerol / Q000494:pharmacology*; D006820:Hyaluronic Acid / Q000494:pharmacology*; D008297:Male; D051379:Mice / Q000196:embryology*; D009028:Morula / Q000187:drug effects*; D020442:Poloxamer / Q000494:pharmacology*; D012710:Serum Albumin, Bovine / Q000494:pharmacology*; D013395:Sucrose / Q000494:pharmacology
D003451:Cryoprotective Agents; D020442:Poloxamer; D012710:Serum Albumin, Bovine; D013395:Sucrose; D006820:Hyaluronic Acid; D005990:Glycerol