Nonsurgical Embryo Transfer Protocol for Use with the NSET™ Device

  • Barbara J. StoneEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2066)


Genetically modified embryos must be transferred to a suitable female recipient for development to pups. Nonsurgical embryo transfer is a fast and efficient method used to deliver blastocyst stage embryos to the uterine horn of recipient females. The efficiency of recovery of pups after nonsurgical embryo transfer is similar to the efficiency after surgical transfer. However, nonsurgical transfer eliminates the pain and distress caused by the surgical procedure and provides a refinement in accordance with Russel and Burch’s “3Rs” (The principles of humane experimental technique. Methuen & Co., London, 1959), an ethical framework for animal research. This method is also useful for rederivation of mouse strains. Rederivation is important for either removal of potential pathogens from an incoming mouse strain after shipping, or within a facility to obtain a clean mouse colony.

Key words

Nonsurgical Embryo Embryo transfer NSET Anesthesia 3Rs Rederivation Transgenic Animal welfare 



Research reported in this publication was funded by the Office of the Director, National Institutes of Health, United States Department of Health and Human Services, under Award Numbers 1R43RR025737-01A1, 2R44RR025737-02, and 8R44OD010958. Additional support was provided by the Kentucky Cabinet for Economic Development, Office of Commercialization and Innovation under the grant agreements KSTC-184-512-11-115 and 184-512-10-096 with the Kentucky Science and Technology Corporation.


  1. 1.
    Russell WMS, Burch RL (1959) The principles of humane experimental technique. Methuen & Co., LondonGoogle Scholar
  2. 2.
    Green M, Bass S, Spear B (2009) A device for the simple and rapid transcervical transfer of mouse embryos eliminates the need for surgery and potential post-operative complications. BioTechniques 47(5):919–924CrossRefGoogle Scholar
  3. 3.
    Steele KH, Hester JM, Stone BJ, Carrico KM, Spear BT, Fath-Goodin A (2013) Nonsurgical embryo transfer device compared with surgery for embryo transfer in mice. J Am Assoc Lab Anim Sci 52(1):17–21PubMedPubMedCentralGoogle Scholar
  4. 4.
    Mainigi MA, Olalere D, Burd I, Sapienza C, Bartolomei M, Coutifaris C (2014) Peri-implantation hormonal milieu: elucidating mechanisms of abnormal placentation and fetal growth. Biol Reprod 90(2):26CrossRefGoogle Scholar
  5. 5.
    de Waal E, Mak W, Calhoun S, Stein P, Ord T, Krapp C, Coutifaris C, Schultz RM, Bartolomei MS (2014) In vitro culture increases the frequency of stochastic epigenetic errors at imprinted genes in placental tissues from mouse Concepti produced through assisted reproductive technologies. Biol Reprod 90(2):22PubMedGoogle Scholar
  6. 6.
    Tian X, Anthony K, Neuberger T, Diaz FJ (2014) Preconception zinc deficiency disrupts postimplantation fetal and placental development in mice. Biol Reprod 90(4):83CrossRefGoogle Scholar
  7. 7.
    Kaufman MR, Albers RE, Keoni C, Kulkarni-Datar K, Natale DR, Brown TL (2014) Important aspects of placental-specific gene transfer. Theriogenology 82(7):1043–1048CrossRefGoogle Scholar
  8. 8.
    Jimenez R, Melo EO, Davydenko O, Ma J, Mainigi M, Franke V, Schultz RM (2015) Maternal SIN3A regulates reprogramming of gene expression during mouse preimplantation development. Biol Reprod 93(4):89CrossRefGoogle Scholar
  9. 9.
    Navarrete FA, Alvau A, Lee HC, Levin LR, Buck J, Martin-De Leon P, Santi CM, Krapf D, Mager J, Fissore RA, Salicioni AM, Darszon A, Visconti PE (2016) Transient exposure to calcium ionophore enables in vitro fertilization in sterile mouse models. Sci Rep 6:33589CrossRefGoogle Scholar
  10. 10.
    Karimi H, Mahdavi P, Fakhari S, Faryabi MR, Esmaeili P, Banafshi O, Mohammadi E, Fathi F, Mokarizadeh A (2017) Altered helper T cell-mediated immune responses in male mice conceived through in vitro fertilization. Reprod Toxicol 69:196–203CrossRefGoogle Scholar
  11. 11.
    Woodford C (2011) Use of a non-surgical embryo transfer (NSET) device as an alternative to rodent surgical embryo transfer (ET) and caesarian re-derivation. Anim Technol Welfare 10(1):42–43Google Scholar
  12. 12.
    Ali RB, van der Ahé F, Braumuller TM, Pritchard C, Krimpenfort P, Berns A, Huijbers IJ (2014) Improved pregnancy and birth rates with routine application of nonsurgical embryo transfer. Trans Res 23(4):691–695CrossRefGoogle Scholar
  13. 13.
    Moreno-Moya JM, Ramírez L, Vilella F, Martínez S, Quinonero A, Noguera I, Pellicer A, Simon C (2014) Complete method to obtain, culture, and transfer mouse blastocysts nonsurgically to study implantation and development. Fertil Steril 101(3):e13CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.ParaTechs CorporationLexingtonUSA

Personalised recommendations