Science et technologie vétérinaires

Cryopreservation of Two Cells Stage Mouse Embryos: An Experience in Center of Molecular Immunology, Cuba

Abstract

González MA, Rodríguez A, Raymond J and González A

Over the past three decades, the generation of genetically modified tumor models has increased. Embryo cryopreservation is essential technique to save and handle the increasing the mice with spontaneous or induced mutations. This technology serves to reduce maintenance costs and the mouse colony is protected from infections. The Center of Molecular Immunology (CIM, Havana, Cuba) have developed several immunotherapeutic projects with focus in oncology studies in some special biomodels. However, there is no experience in the field of cryopreservation methods as a tool to increase the quality to work with genetically engineered mouse models. The objective of this work is to establish the technique of cryopreservation of murine embryos at the CIM. The maintenance and handling of animals from the National Laboratory Animal Production Center (CENPALAB), was carried under the regulations of the Institutional Committee for the Care and Use of Laboratory Animals (CICUAL) of the CIM. We used a vitrification method for mouse embryos in liquid nitrogen with etylenglycol medium. In addition, a Ficoll and sucrose was employed to prevent the devitrification process. The freeze embryos since four months were transferred to the pseudopregnant (NMRI/Cenp mice). The efficiency of the defrosting process was greater than 80%. As a result of the embryo transfer, the healthy born pups were obtained. These results suggest that the cryopreservation technique was established at the CIM and it will promote high standards of biomedical research.