Freezing of in Vivo Derived and in Vitro Pre-Cultured Porcine Blastocysts : Differences of the Cryoprotective Effect between Glycerol and Ethylene Glycol

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The present study was conducted to compare glycerol and ethylene glycol as cryoprotectants for porcine in vivo derived and in vitro pre-cultured peri-hatching blastocysts. Embryos were surgically collected from superovulated gilts on Day 6. In vivo derived hatched blastocysts (in vivo HB) were frozen with 1.5 M glycerol (1.5 G) or 1.5 M ethylene glycol (1.5 EG). In vitro pre-cultured peri-hatching blastocysts (in vitro pHB), obtained by culturing Day 6 embryos for 18 h in Whittens medium supplemented with 1.5% bovine serum albumin and with or without 5% fetal calf serum (FCS), were frozen with 1.5 G or 1.8 EG. All embryos were frozen by a slow cooling method using a programmable freezer. The survival of in vivo HB frozen with 1.5 G (82%) was higher (P<0.01) compared with embryos frozen with 1.5 EG (36%). More blastocysts cultured with FCS developed to the hatched blastocyst stage compared with those cultured without FCS (73% vs 33%, P<0.01) within 18 h. The survival of embryos pre-cultured with or without FCS however was not different following freezing with 1.5 G (23% vs 34%) or 1.8 EG (74% vs 81%). The survival rates for embryos frozen with 1.8 EG were significantly higher (P<0.01) than those for the 1.5 G groups. These results demonstrate that more than 80% of in vivo HB can be successfully frozen with 1.5 G and that similar survival rates for in vitro pHB can be achieved by using 1.8 EG. The study also indicates inherent differences in the post-thaw survival of in vivo derived and in vitro pre-cultured peri-hatching blastocysts frozen with 1.5 G.
日本繁殖生物学会の論文
1996-08-01

Freezing of in Vivo Derived and in Vitro Pre-Cultured Porcine Blastocysts : Differences of the Cryoprotective Effect between Glycerol and Ethylene Glycol

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