Overview of new developments in and the future of cryopreservation in the laboratory mouse
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Overview of new developments in and the future of cryopreservation in the laboratory mouse Mo Guan • Susan Marschall • Marcello Raspa Amanda R. Pickard • Martin D. Fray
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Received: 10 April 2012 / Accepted: 30 July 2012 / Published online: 31 August 2012 Ó Springer Science+Business Media, LLC 2012
Introduction to cryopreservation
freezing embryos at -196 °C (Whittingham et al. 1972; Wilmut 1972). Since then the field has developed to include a wide range of procedures that can be exploited to archive as well as reanimate mouse stocks from frozen embryos, gametes, and ovarian tissues (Fray 2009a, b). Embryos, gametes, and ovarian tissue frozen and handled in an appropriate manner will remain viable indefinitely at liquid nitrogen (LN2) temperature because all biological reactions effectively stop below the glass transition temperature of water (-130 °C). This statement is supported by data that show that frozen embryos exposed to radiation levels equivalent to 2,000 years (200 cGy) of background radiation develop normally post thawing (Glenister et al. 1984). Live animals have also been recovered from spermatozoa thawed out after being cryopreserved for many years (Kaneko et al. 2006; Leibo et al. 1994).
Cryopreserving the laboratory mouse came of age in the early 1970s with the publication of robust protocols for
Application and limitations of embryo freezing
Abstract The large-scale mutagenesis programmes underway around the world are generating thousands of novel GA mouse strains that need to be securely archived. In parallel with advances in mutagenesis, the procedures used to cryopreserve mouse stocks are being continually refined in order to keep pace with demand. Moreover, the construction of extensive research infrastructures for systematic phenotyping is fuelling demand for these novel strains of mice and new approaches to the distribution of frozen and unfrozen embryos and gametes are being developed in order to reduce the dependency on the transportation of live mice. This article highlights some contemporary techniques used to archive, rederive, and transport mouse strains around the world.
M. Guan A. R. Pickard M. D. Fray (&) Mary Lyon Centre, Medical Research Council, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, UK e-mail: [email protected] S. Marschall Institute of Experimental Genetics, Helmholtz Zentrum Mu¨nchen, German Research Center for Environmental Health (GmbH), Ingolsta¨dter Landstrasse 1, 85764 Neuherberg, Germany M. Raspa Istituto di Biologia Cellulare e Neurobiologia, Consiglio Nazionale delle Ricerche (CNR-EMMA), Campus ‘‘Adriano Buzzati-Traverso’’, Via E. Ramarini 32, 00015 Monterotondo Scalo, Italy
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Embryo freezing has traditionally been the preferred method for archiving mouse lines, and many protocols using a variety of cryoprotectants, e.g., propylene glycol, ethylene glycol, and dimethylsulphoxide (DMSO), have been adopted successfully. The objective is to recover sufficient live-born offspring to reestablish a breeding colony from a single straw/vi
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