Willadsen S, Polge C, Rowson L E
J Reprod Fertil. 1978 Mar;52(2):391-3. doi: 10.1530/jrf.0.0520391.
Day 7 cow embryos were frozen in 1.5 M-DMSO in PBS at 0.3 degrees C/min to -36 degrees C and at 0.1 degrees C/min between -36 and -60 degrees C before being plunged directly into liquid nitrogen. They were subsequently thawed (rapidly to -50 degrees C, at 4 degrees C/min from -50 to -10 degrees C, and rapidly again) to room temperature. Embryonic viability was tested by four different transfer techniques. Maximum pregnancy rate (8/12) was obtained with surgical transfer immediately after thawing and dilution of DMSO.
将第7天的奶牛胚胎在含1.5M二甲基亚砜(DMSO)的磷酸盐缓冲液(PBS)中,以0.3℃/分钟的速度降温至-36℃,然后在-36℃至-60℃之间以0.1℃/分钟的速度降温,之后直接投入液氮中。随后将它们解冻(快速升温至-50℃,从-50℃以4℃/分钟的速度升温至-10℃,然后再次快速升温)至室温。通过四种不同的移植技术检测胚胎活力。解冻并稀释DMSO后立即进行手术移植,获得了最高妊娠率(8/12)。