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Open Access Cryopreservation Methodologies: Freezing Location and Protocol Alter Development of Mouse Embryos

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The purpose of this study was to determine the effects of location in the freezing chamber and various freezing protocols on post-thaw development of two-cell mouse embryos. Different protocols were used to cryopreserve two-cell mouse embryos in different freezer positions. Embryos were thawed and cultured to observe post-thaw development. When four different freezer positions were compared, one position provided better post-thaw blastocyst development, regardless of several protocols tested. Furthermore, there were significant differences in temperatures between positions in the freezer. These differences came before seeding (–7° C), just before changing the –7.0° C hold demand (Ramp 2), during the two time points tested during Ramp 3 (–0.3° C/min), and during the holding ramp of –30° C. Post-thaw embryo development was improved by lengthening the time after seeding but before decreasing the temperature again. Decreasing embryos' temperature before plunging appeared to improve post-thaw performance.

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Document Type: Case Report

Affiliations: 1: Department of Obstetrics and Gynecology, Greenville Hospital System, Greenville, South Carolina 2: Department of Quality Management, Greenville Hospital System, Greenville, South Carolina

Publication date: January 1, 2004

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  • The Journal of the American Association for Laboratory Animal Science (JAALAS) serves as an official communication vehicle for the American Association for Laboratory Animal Science (AALAS). The journal includes a section of refereed articles and a section of AALAS association news. The mission of the refereed section of the journal is to disseminate high-quality, peer-reviewed information on animal biology, technology, facility operations, management, and compliance as relevant to the AALAS membership. JAALAS accepts research reports (data-based) or scholarly reports (literature-based), with the caveat that all articles, including solicited manuscripts, must include appropriate references and must undergo peer review.

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