Efficiency of timed pregnancies in C57BL/6 and BALB/c mice by mating one male with up to four females.

For a wide range of biomedical approaches, an accurate estimate of the age of embryos or pups is important. Overnight mating is the method that is mostly used to establish timed pregnancies. The oestrus cycle in mice repeats every four to five days. So, not all females will get pregnant because they are not in oestrus. Therefore, the aim of this study was to analyse whether polygamous mating could increase the rate of timed pregnancies per breeding cage and female. We compared overnight timed mating regimes with up to four females per male, using C57BL/6 and BALB/c mice as well as F1 hybrids of these two strains. The number of vaginal plugs, number of females that gave birth and weaned litter (including size and weaning weight) were recorded. Our results showed that the plug and pregnancy rate decreased, but the productivity per breeding cage increased for polygamous mating regimes. The proportion of females with vaginal plugs and females that gave birth was significantly higher in monogamous mating. The proportion of plugged females that gave birth, as well as litter size and weaning weight, were not influenced by the mating regime. After analysing 513 breeding cages with a total of 1090 females, we found that polygamous mating with up to three females per male can increase the number of timed pregnancies. However, in the mating regime with more than three females, the rate of timed pregnancy as well as number of pups per female declined.

Success of embryo transfer in mice with freshly collected and cryopreserved two-cell embryos with different genetic backgrounds correlated with the number of transferred embryos: A 5-year retrospective analysis.

Embryo transfer of pre-implantation embryos to surrogate dams is a key technique for the hygienic sanitation of strains, cryopreservation, in vitro fertilization, genetic modification and engineering. However, the effects of several parameters, such as the number of transferred embryos, on the success of embryo transfer are not well studied. In this retrospective study, we reanalysed 1320 embryo transfers of two-cell embryos originating from genetically altered donors, which were performed under routine conditions in our facility over a period of 5 years. Of them, 453 embryo transfers were done with freshly collected embryos and 867 transfers were performed with cryopreserved embryos. Despite the fact that the genetic background of the embryo donors was quite heterogeneous, we found that the transfer of ≥ 21 embryos reduced the success of embryo transfers for freshly collected embryos in correlation with the number of pregnancies and born pups, whereas this was not the case for transfer in the cryopreservation group. Most pregnancies were achieved after embryo transfer of 10-20 freshly collected embryos (90.4%), which dropped to 37.5% if more embryos were transferred. The highest pregnancy rates in the cryopreservation group were achieved if 15-17 embryos were transferred (62.9%). Despite the fact that the precise substrains were only rarely defined, we confirmed that beside the number of transferred embryos, the genetic background of the donors had an influence on the success of embryo transfer. Significantly more embryos in a C57BL/6 background developed to term than embryos on a BALB/c background.

Direct comparison of vasectomized males and genetically sterile Gapdhs knockout males for the induction of pseudopregnancy in mice.

The laboratory mouse is the most used animal model in biomedical research. Several artificial reproductive techniques, such as revitalization of cryopreserved strains, rederivation after hygienic contaminations and the production of transgenic mouse models, require the transfer of preimplantation embryos to surrogate mothers. Pseudopregnancy is essential in recipient females and is induced by mating with sterile males. Commonly, surgically vasectomized males are used for this purpose. As an alternative, genetically modified mouse strains have been identified, in which homozygous infertile males are sexually active. Here, we investigated the suitability of genetically infertile Gapdhstm1Dao males under routine laboratory conditions with respect to plug rates, pregnancy rates and frequency of born offspring after embryo transfer. Our results showed no significant differences for these aspects between Gapdhstm1Dao and vasectomized CD2F1 males. In addition, we evaluated the efforts to obtain a defined number of sterile males either by breeding of sterile mutants or surgical vasectomy, and addressed the impact of both options on animal welfare. In conclusion, infertile males of the Gapdhstm1Dao line are a reliable alternative to vasectomized males for the induction of pseudopregnancy, and can contribute to the refinement of the procedure by avoiding surgical interventions.