5. ガラス化溶液への種々の単糖類の添加による牛体外受精由来胚の生存性への影響(セミナー「ビトリフィケーション」)

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The effect of the addition of various monosaccharides to a vitrification solution on the survival rate of bovine blastocysts produced in vitro was investigated. In vitro matured and in vitro fertilized bovine Day 7-8 blastocysts were cassified into three developmental stages:early blastocysts (EB), blastocysts (BL), and expanded blastocysts (exB). Blastocysts were cryopreserved in one of the vitrification solutions which consists of 20% glycerol + 20% ethylene glycol + 0.3M sucrose + 3% polyethylene glycol + 0.3M (monosaccharide). As a monosaccharide, fructose, glucose, inositol, mannitol, sorbitol and xylose were respectively employed in the vitrification solution above. The basic solution was Dulbecco's PBS. Embryos were exposed to each vitrification solution in three steps, and after loading to 0.25ml straws, were plunged into liquid nitrogen. After warming in water bath at 20℃, cryoprotectants were diluted in 1/2 and 1/4M sucrose solution each for 5 min. Equilibration and dilution procedures except warming were conducted at room temperature (23 to 27℃). After dilution, embryos were cultured in Ham's F10 + 0.1mM β-mercaptoethanol + 20% fatal calf serum. Survival rates at 48 hr. (hatching rate at 96hr.) incubation of all stages of blastocysts exposed to six types of the monosaccharide additions and vitrified were 65.6 (55.5), 69.6 (56.3), 71.3 (59.1), 66.1 (56.7), 66.7 (56.7), 85.2 (79.3) % in sorbitol, mannitol, fructose, glucose, inositol and xylose respectively. Xylose-addition to the vitrification solution showed significantly higher survival rate in both 48 hr. survival rate (P<0.05) and 96 hr. hatching rate (P<0.001) than 5 other monosaccharrides addition. Within every solutions, more developed staged blastocysts showed higher survival rate. These results indicate that the addition of xylose to the vitrification solution improves the survival of vitrified bovine embryos produced in vitro.
低温生物工学会の論文
1997-07-11

5. ガラス化溶液への種々の単糖類の添加による牛体外受精由来胚の生存性への影響(セミナー「ビトリフィケーション」)

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