Horvath G, Seidel G E
Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins, CO, USASZIE, Faculty of Veterinary Science, Department of Obstetrics and Reproduction, Budapest, Hungary.
Reprod Domest Anim. 2008 Jun;43(3):333-338. doi: 10.1111/j.1439-0531.2007.00905.x. Epub 2007 Dec 7.
After vitrification of oocytes, fertilization rates and subsequent development are unsatisfactory, possibly due in part to zona hardening. Foetal calf serum (FCS) can prevent zona hardening because of its fetuin content, but FCS composition varies among batches, and may contain viruses. In this study, we therefore compared media supplemented with different sources of macromolecules, 2% bovine serum albumin (BSA), 2% BSA + 1 mg/ml fetuin and 20% FCS, for handling oocytes for 10-30 min prior to vitrification. None of the treatments resulted in developmental rates comparable with the non-vitrified controls, but FCS inclusion in pre-vitrification handling medium resulted in higher blastocyst production per oocyte (p < 0.05) (10.8%) on day 9 of culture than BSA (5.3%) or BSA + fetuin (6.4%). Blastocysts developing from oocytes from all vitrification treatments were somewhat retarded relative to those developed from non-vitrified oocytes. We also tested the use of fetuin during vitrification as well as two different exposure times with cryoprotectants, 180 and 30 s. There was no significant effect of fetuin or exposure time on rates of subsequent blastocyst production.
卵母细胞玻璃化冷冻后,受精率和后续发育情况并不理想,部分原因可能是透明带硬化。胎牛血清(FCS)因其含有胎球蛋白可防止透明带硬化,但不同批次的FCS成分存在差异,且可能含有病毒。因此,在本研究中,我们比较了在卵母细胞玻璃化冷冻前用不同大分子来源进行处理的培养基,即2%牛血清白蛋白(BSA)、2% BSA + 1 mg/ml胎球蛋白和20% FCS,处理时间为10 - 30分钟。所有处理组的发育率均不及未冷冻对照组,但在玻璃化冷冻前处理培养基中添加FCS,在培养第9天时,每个卵母细胞产生的囊胚率(10.8%)高于添加BSA组(5.3%)或BSA +胎球蛋白组(6.4%)(p < 0.05)。相对于未玻璃化冷冻的卵母细胞发育形成的囊胚,所有玻璃化冷冻处理的卵母细胞发育形成的囊胚发育均有所延迟。我们还测试了在玻璃化冷冻过程中使用胎球蛋白以及两种不同的冷冻保护剂暴露时间,即180秒和30秒。胎球蛋白或暴露时间对后续囊胚产生率均无显著影响。