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KMID : 0900919930170030201
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Korean journal of Animal Reproduction
1993 Volume.17 No. 3 p.201 ~ p.207
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Ultrarapid Freezing of DNA-Injected and Nuclear-transplanted Mouse Embryos
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Han Yong-Mahn
Lee Chul-Sang
Yu Dae-Yeul
Shin Sang-Tae
Lee Kyung-Kwang
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Abstract
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We determined whether the ultrarapid freezing method is applicable to micromanipulated mouse embryos. One-cell mouse embryos were microinjected with MThGH gene. Nuclei from one-cell embryos of F1(C57BLtimesCBA) mice were transplanted into enucleated one-cell embryos of ICR mice. The injected and nucleated embryos that developed to 2-cell stage were cryopreserved by ultrarapidfreezing. The embryos equilibrated in freezing medium(3 M DMSO＋0.25 M sucrose＋2% FBS in PBS) were directly immersed into liquid nitrogen and then thawed in 37^{circ}C water. Development rates of the microinjected and nuclear-transplanted embryos to blastocyst stage after ultrarapidly freezing and thawing were 31% and 55%, respectively. The frozen-thawed embryos were transferred to pseudopregnant recipient, which then gave birth to 17 offsprings. Twelve(14% of the transferred embryos) and five(20%) offsprings were derived from microinjected and nuclear-transplanted embryos, respectively. The results indicate that the DNA injected and nuclear-transplanted mouse embryos are cryopreservable at 2-cell stage by ultrarapid freezing method.
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