Cryopreservation of mouse half-morulae and chimeric embryos by vitrification.
Mouse half-morulae were cryopreserved less than or equal to 1, 3, 6, and 12 hr after bisection by the vitrification method using 25% glycerol and 25% 1,2-propanediol as cryoprotectant. The developmental rates of the frozen-thawed half-embryos to blastocysts in vitro were 77.8% (63/81), 82.0% (41/50), 92.1% (117/127), and 0% (0/37), respectively. Sixty-one of the half-embryos that had been vitrified 6 hr after the bisection followed by transfer to five recipients resulted in a total of ten (16.4%) normal fetuses. Chimeric mouse embryos constructed by two half-morulae were also vitrified 6 and 16 hr after aggregation. Survivors were obtained from the former case: 40 (80.0%) of 50 frozen-thawed embryos developed in vitro to blastocysts, and, after transfer, six chimeric offspring were obtained from the 34 vitrified chimeric embryos. These results showed that mouse half-morulae and chimeric embryos could be cryopreserved by the vitrification method. It seems possible to manufacture a chimeric mouse embryo of defined genotypic composition that can be analyzed during its frozen state using the identical half-embryos of the components.
Nagashima
H
H
Research Center, Nisshin Flour Milling Co., Ltd., Tochigi, Japan.
Kobayashi
K
K
Yamakawa
H
H
Ogawa
S
S
D000818:Animals; D002678:Chimera / Q000502:physiology*; D015925:Cryopreservation / Q000379:methods*; D004624:Embryo Transfer; D005260:Female; D051379:Mice; D008815:Mice, Inbred Strains; D008866:Microsurgery; D009028:Morula / Q000502:physiology / Q000637:transplantation*; D011247:Pregnancy; D014022:Tissue Survival / Q000502:physiology