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Open Access Cryopreservation of Murine Zygotes for Use in Testing Culture Environments

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Developing one-cell mouse zygotes are more sensitive to in vitro environmental conditions than are cleavage-stage embryos. However, for convenience and reproducibility, cryopreserved two-cell zygotes are routinely used for such assays. Concern over the possibility of inducing damage by exposing one-cell zygotes to cryoprotective agents and freeze-thaw procedures during syngamy led us to examine one-cell zygotes, with and without visible pronuclei, in an effort to minimize or avoid these effects and obtain the highest possible developmental rate. In vivo fertilized mouse zygotes were collected 21 to 43 h after administration of human chorionic gonadotropin (hCG). Suspensions of zygotes in 2M ethylene glycol were aspirated into 0.25-ml plastic insemination straws and slowly cooled at -0.5°C/min to -40°C before being plunged into liquid nitrogen for storage. Zygotes were thawed, rinsed, and placed in culture. Zygotes were examined initially for damage from the freeze-thaw procedure. Daily in vitro development was recorded. In this group of zygotes, no damage was apparent immediately after thawing, and a high degree of development in vitro was observed. Thus, usefulness of a cryopreservation method for one-cell murine zygotes has been confirmed.

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Document Type: Research Article

Publication date: October 1, 1997

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  • Comparative Medicine (CM), an international journal of comparative and experimental medicine, is the leading English-language publication in the field and is ranked by the Science Citation Index in the upper third of all scientific journals. The mission of CM is to disseminate high-quality, peer-reviewed information that expands biomedical knowledge and promotes human and animal health through the study of laboratory animal disease, animal models of disease, and basic biologic mechanisms related to disease in people and animals.

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