Skip to main content

Nonsurgical Embryo Transfer Protocol for Use with the NSET™ Device

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

This is a preview of subscription content, access via your institution.

Buying options

USD   49.95
Price excludes VAT (USA)
  • DOI: 10.1007/978-1-4939-9837-1_9
  • Chapter length: 5 pages
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
USD   109.00
Price excludes VAT (USA)
  • ISBN: 978-1-4939-9837-1
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
Softcover Book
USD   149.99
Price excludes VAT (USA)
Hardcover Book
USD   219.99
Price excludes VAT (USA)
Fig. 1

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more


  1. Russell WMS, Burch RL (1959) The principles of humane experimental technique. Methuen & Co., London

    Google Scholar 

  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):919–924

    CAS  CrossRef  Google Scholar 

  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):17–21

    CAS  PubMed  PubMed Central  Google Scholar 

  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):26

    CrossRef  Google Scholar 

  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):22

    PubMed  Google Scholar 

  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):83

    CrossRef  Google Scholar 

  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):1043–1048

    CAS  CrossRef  Google Scholar 

  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):89

    CrossRef  Google Scholar 

  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 6:33589

    CAS  CrossRef  Google Scholar 

  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 69:196–203

    CAS  CrossRef  Google Scholar 

  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):42–43

    Google Scholar 

  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):691–695

    CrossRef  Google Scholar 

  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):e13

    CrossRef  Google Scholar 

Download references


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.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Barbara J. Stone .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and Permissions

Copyright information

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

About this protocol

Verify currency and authenticity via CrossMark

Cite this protocol

Stone, B.J. (2020). Nonsurgical Embryo Transfer Protocol for Use with the NSET™ Device. In: Larson, M. (eds) Transgenic Mouse. Methods in Molecular Biology, vol 2066. Humana, New York, NY.

Download citation

  • DOI:

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9836-4

  • Online ISBN: 978-1-4939-9837-1

  • eBook Packages: Springer Protocols