Leibo S P, DeMayo F J, O'Malley B
Department of Obstetrics & Gynecology, Baylor College of Medicine, Houston, Texas.
Mol Reprod Dev. 1991 Dec;30(4):313-9. doi: 10.1002/mrd.1080300405.
Cryopreserved fertilized mouse ova were used to generate transgenic mice via micromanipulation. Five-DNA constructions were injected into a total of 1,052 cryopreserved ova, of which 683 (65%) survived the injection and were transferred into recipients. Of 35 recipients, 66% became pregnant and littered a total of 88 pups. As controls, these DNA constructions were also injected into 1,123 fresh ova, of which 744 (66%) survived and were transferred. Of 42 recipients, 79% became pregnant and littered a total of 167 pups. That is, 22% of fresh ova that were transferred developed into live pups, whereas only 13% of cryopreserved ova did so. Of the pups born, 42 of the 167 (25%) produced from fresh ova were transgenic, and 28 of the 88 (32%) produced from cryopreserved ova were transgenic. In terms of the injected ova that had been transferred, 5.6% of the 744 fresh and 4.1% of the 683 frozen ova developed into transgenic mice. These data indicate that the efficiency of production of transgenic mice from cryopreserved ova is close to that from fresh ova. That observation and the fact that cryopreserved ova allow more efficient utilization of animals suggest that cryopreserved ova can be used instead of fresh ova to produce transgenic mice.
冷冻保存的受精小鼠卵子通过显微操作用于生成转基因小鼠。将五种DNA构建体注射到总共1052个冷冻保存的卵子中，其中683个（65%）在注射后存活并被移植到受体中。在35个受体中，66%怀孕并总共产下88只幼崽。作为对照，这些DNA构建体也被注射到1123个新鲜卵子中，其中744个（66%）存活并被移植。在42个受体中，79%怀孕并总共产下167只幼崽。也就是说，移植的新鲜卵子中有22%发育成活幼崽，而冷冻保存的卵子中只有13%如此。在出生的幼崽中，167只由新鲜卵子产生的幼崽中有42只（25%）是转基因的，88只由冷冻保存的卵子产生的幼崽中有28只（32%）是转基因的。就已移植的注射卵子而言，744个新鲜卵子中有5.6%以及683个冷冻卵子中有4.1%发育成转基因小鼠。这些数据表明，从冷冻保存的卵子生产转基因小鼠的效率与从新鲜卵子生产的效率相近。这一观察结果以及冷冻保存的卵子能更高效利用动物这一事实表明，冷冻保存的卵子可替代新鲜卵子用于生产转基因小鼠。
