Complete method to obtain culture and transfer mouse blastocysts nonsurgically to study implantation and development

Original Video Article

Complete method to obtain, culture, and transfer mouse blastocysts nonsurgically to study implantation and development

Authors:
Juan Manuel Moreno-Moya, M.Sc., Leslie B. Ramírez, M.Sc., Felipe Vilella, Ph.D., Sebastián Martínez, B.Sc., Alicia Quiñonero, B.Sc., Inmaculada Noguera, Ph.D., Antonio Pellicer, M.D., Ph.D., Carlos Simón, M.D., Ph.D.

Capsule:
This step-by-step video explains how to obtain, culture, and transfer blastocysts in order to study implantation and development using a new non-surgical method.

Objective:
This video article details an efficient and complete step-by-step protocol for studying implantation in mice.
Design:
A video presentation of an animal model for research in reproductive biology.
Setting:
Pseudopregnant mice; mouse embryo production, recovery, culture, and transfer.
Animal(s):
Mouse (Mus musculus).
Intervention(s):
Implantation is an essential step in human reproduction although, because of technical and ethical considerations, still relatively little is known about human implantation and early development. Conversely, mouse models are wellestablished and can be used for preliminary experiments. However, there are various bottlenecks in the procedure for obtaining and transferring murine embryos, which make experimentation with this model more difficult. These difficulties include: pseudopregnancy, ovarian hyperstimulation, and embryo collection, culture, and transfer. We propose here a complete and efficient method for obtaining, culturing, and transferring mouse blastocysts that can be easily applied in research. Applications might include testing new media components which do not affect pre-implantation but do affect implantation and early development. We propose using a non-surgical embryo transfer system which is very similar to the one used for human embryo transfer (1-5).
Main Outcome Measure(s):
All animal experiments were carried out with approval from the Institutional Review Board. In this video the protocols with recipient and donor mice were carried out in parallel in the same week. For the donor mice: the first step is ovarian stimulation, followed by ovulation induction and then mating; finally the mice are sacrificed and the embryos are collected and cultured. For recipient mice: firstly estrous synchrony is induced, followed by mating with a vasectomized male, next the vaginal plug is visualized, and the embryos are transferred non-surgically. Finally (optionally), the implantation sites can be visualized on day 7.5 of development.
Results:
The embryo transfer method proposed here has demonstrated previously to achieve embryo implantation easier, faster and in approximately similar rates than another traditional surgery methods.
Conclusion(s):
This workflow is the first set of complete step-by-step instructions available which incorporates advances such as non-surgical mouse embryo transfer. This will facilitate research into different reproduction events like embryo development, embryo implantation, or contraception.
References
1. Deb K, Reese J, Paria BC. Methodologies to study implantation in mice. Methods Mol Med 2006;121:9-34.
2. L. F. van Zutphen, V. Baumans, A. C. Beynen. Principles of Laboratory Animal Science, Revised Edition. May 2001.
3. Shea K, Geijsen N. Dissection of 6.5 dpc mouse embryos. J Vis Exp 2007;(2):160. doi:160.
4. Steele KH, Hester JM, Stone BJ, Carrico KM, Spear BT, Fath-Goodin A. Nonsurgical embryo transfer device compared with surgery for embryo transfer in mice. J Am Assoc Lab Anim Sci 2013;52:17-21.

 

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