Nguyen Van Khanh, Vu Huong Thi Thu, Nguyen Huong Thi, Quan Huu Xuan, Pham Lan Doan, Kikuchi Kazuhiro, Nguyen Son Thanh, Somfai Tamas
Key Lab of Animal Cell Biotechnology, National Institute of Animal Science, Hanoi, Vietnam.
Division of Animal Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Ibaraki 305-8602, Japan.
J Reprod Dev. 2018 Oct 12;64(5):457-462. doi: 10.1262/jrd.2018-047. Epub 2018 Aug 10.
We compared the efficacy of the microdrop and minimum volume cooling (MVC) methods for the vitrification of in vitro-produced porcine zygotes and blastocysts after equilibration in low concentrations of cryoprotectant agents. Zygotes and blastocysts were equilibrated in 2% (v/v) ethylene glycol and 2% (v/v) propylene glycol for 13-15 min. Then, they were vitrified in a medium comprised of 17.5% ethylene glycol, 17.5% propylene glycol, 0.3 M sucrose, and 50 mg/ml polyvinylpyrrolidone either by either dropping them directly into liquid nitrogen (microdrop method) or placing them on Cryotop sheets in a minimum volume of medium and plunging into liquid nitrogen (MVC method). Both zygotes and blastocysts were successfully vitrified. For the vitrification of zygotes, the MVC and microdrop methods were equally effective; however, for blastocyst vitrification, MVC was superior. For both methods, the vitrification of zygotes produced higher-quality embryos than the vitrification of blastocysts.
我们比较了在低浓度冷冻保护剂中平衡后，微滴法和最小体积冷却（MVC）法对体外生产的猪受精卵和囊胚进行玻璃化冷冻的效果。将受精卵和囊胚在2%（v/v）乙二醇和2%（v/v）丙二醇中平衡13 - 15分钟。然后，将它们置于由17.5%乙二醇、17.5%丙二醇、0.3 M蔗糖和50 mg/ml聚乙烯吡咯烷酮组成的培养基中进行玻璃化冷冻，方法是直接将其滴入液氮中（微滴法），或者将其置于最小体积培养基中的Cryotop载片上并投入液氮中（MVC法）。受精卵和囊胚均成功实现玻璃化冷冻。对于受精卵的玻璃化冷冻，MVC法和微滴法效果相同；然而，对于囊胚的玻璃化冷冻，MVC法更具优势。对于这两种方法，受精卵的玻璃化冷冻产生的胚胎质量高于囊胚的玻璃化冷冻。
