Animal Biotechnology Centre, Nanaji Deshmukh Veterinary Science University, Jabalpur, Madhya Pradesh, India.
Cryo Letters. 2021 Jan-Feb;42(1):13-18.
Vitrification is an ultra-rapid freezing technique for germplasm preservation under high salt concentration with very short exposure time.
To assess the post-thawed developmental potential of in vitro-produced buffalo embryos vitrified by solid surface technique using different concentrations of cryoprotectants.
The slaughterhouse derived oocytes were in vitro matured and fertilized with epididymal sperm. IVF embryos at the morula stage were vitrified under two protocols; (i) Protocol-1: ethylene glycol (35%) (ii) Protocol-2: ethylene glycol (15%) and dimethyl sulfoxide (15%). The vitrified-thawed embryos were in vitro cultured up to the blastocyst stage.
Post-thawed development of embryos vitrified under Protocol-1 was significantly higher in terms of compact morula formation as compared to Protocol-2. However, blastocyst developmental rates were not significantly different between the two protocols. The developmental rates of the non-vitrified control were significantly higher than embryos vitrified by either protocols.
The process of cryopreservation, under both protocols, significantly affected the developmental potential of pre-implant embryos as compared to fresh embryos. Hence the nature and concentrations of cryoprotectants needs to be optimized for efficient, viable embryonic development.
玻璃化是一种在高盐浓度下通过极短时间暴露来保存种质的超快速冷冻技术。
评估使用不同浓度的冷冻保护剂通过固相技术对体外生产的水牛胚胎进行玻璃化冷冻后的解冻后发育潜力。
屠宰场来源的卵母细胞进行体外成熟,并与附睾精子受精。处于桑葚胚阶段的 IVF 胚胎根据两种方案进行玻璃化处理;(i)方案 1:乙二醇(35%)(ii)方案 2:乙二醇(15%)和二甲基亚砜(15%)。将玻璃化-解冻的胚胎在体外培养至囊胚阶段。
与方案 2 相比,方案 1 下玻璃化冷冻的胚胎在致密桑葚胚形成方面的解冻后发育明显更高。然而,两种方案之间的囊胚发育率没有显著差异。非玻璃化对照组的发育率明显高于两种方案玻璃化冷冻的胚胎。
与新鲜胚胎相比,两种方案下的冷冻保存过程都显著影响了胚胎的发育潜力。因此,需要优化冷冻保护剂的性质和浓度,以实现高效、可行的胚胎发育。