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Complete method to obtain, culture, and transfer mouse blastocysts nonsurgically to study implantation and
Juan Manuel Moreno-Moya1, Leslie Ramírez1, Felipe Vilella1
1Fundación Instituto Valenciano de Infertilidad (FIVI) and Instituto Universitario IVI/INCLIVA (Investigación Clínico de Valencia), Valencia University, Valencia, Spain.
Objective:
To illustrate an efficient, complete, step-by-step protocol for studying implantation in mice.
Design:
Video presentation of an animal model for research in reproductive biology.
Animal(S):
Mouse (Mus musculus).
Intervention(S):
A nonsurgical embryo transfer system very similar to that used for human embryo transfer.
Main Outcome Measure(S):
The protocols with recipient and donor mice are performed in parallel in the same week. For the donor mice: the first step is ovarian stimulation, followed by ovulation induction and mating; finally, the mice are sacrificed, and the embryos are collected and cultured. For recipient mice: first estrous synchrony is induced, followed by mating with a vasectomized male, visualization of the vaginal plug, and nonsurgical transfer of the embryos. Finally (optionally), the implantation sites can be visualized on day 7.5 of development. (All animal experiments were performed with the approval of the institutional review board.)
Result(S):
Implantation is an essential step in human reproduction although, because of technical and ethics considerations, still relatively little is known about human implantation and early development. Conversely, mouse models are well established and can be used for preliminary experiments. However, there are various bottlenecks in the procedure for obtaining and transferring murine embryos, which makes experimentation with this model more difficult. These difficulties include pseudopregnancy, ovarian hyperstimulation, and embryo collection, culture, and transfer. We have proposed a complete, efficient method for obtaining, culturing, and transferring mouse blastocysts that can be easily applied in research. Potential applications include testing new media components that do not affect preimplantation but do affect implantation and early development. The embryo transfer method proposed here has been demonstrated to achieve embryo implantation easier and faster than, and in approximately similar rates as other traditional surgery methods.
Conclusion(S):
This workflow is the first set of complete step-by-step instructions available that incorporate advances such as nonsurgical mouse embryo transfer. This will facilitate research into different reproduction events such as embryo development, embryo implantation, or contraception.
