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Cryopreservation of mouse half‐morulae and chimeric embryos by vitrification
Author(s) -
Nagashima Hiroshi,
Kobayashi Kazuhiko,
Yamakawa Hirohito,
Ogawa Shyoso
Publication year - 1991
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.1080300309
Subject(s) - vitrification , cryopreservation , cryoprotectant , embryo , andrology , biology , embryo transfer , embryo cryopreservation , genetics , medicine
Mouse half‐morulae were cryopreserved ≤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.