Transgenic Research

, Volume 14, Issue 5, pp 685–690 | Cite as

Cryopreserved Morulae can be used to Efficiently Generate Germline-transmitting Chimeras by Blastocyst Injection

  • Janice V. Parker-ThornburgEmail author
  • Jennifer L. Alana
  • Chad N. Smith
  • Michelle Detry
  • Marta L. Rojas
  • Kedryn K. Baskin
Technical report


The production of chimeric mice is a complex process, requiring the careful coordination of tissue culture cell growth, production of a large number (30–75) of competent blastocysts and the availability of appropriately timed pseudo pregnant female mice. Failure at any of these steps can impinge upon the rapid production of chimeras. One potential improvement for the efficient generation of chimeric mice would be the utilization of cryopreserved embryos suitable for injection. C57Bl/6 morulae were frozen using a standard 2-step protocol with ethylene glycol as the cryopreservation agent. We determined that cryopreserved morulae could thaw, culture to blastocyst stage in KSOM media and survive injection at rates equivalent to control embryos. Cryopreserved morulae were also equivalent to controls at all later stages in the process of production of chimeric mice, including birth rate, percentage chimerism of resulting animals and ability to produce germline progeny. Hence, cryopreservation of morulae for blastocyst injection is a suitable option to enhance the efficiency of chimeric mouse generation.


blastocyst injection cryopreservation embryo freezing ES cells germline transmission 


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Copyright information

© Springer 2005

Authors and Affiliations

  • Janice V. Parker-Thornburg
    • 1
    Email author
  • Jennifer L. Alana
    • 2
  • Chad N. Smith
    • 2
  • Michelle Detry
    • 3
  • Marta L. Rojas
    • 1
  • Kedryn K. Baskin
    • 1
  1. 1.Department of Biochemistry and Molecular BiologyUniversity of Texas M. D. Anderson Cancer Center Genetically Engineered Mouse FacilityHoustonUSA
  2. 2.Department of Molecular GeneticsUniversity of Texas M. D. Anderson Cancer Center Genetically Engineered Mouse FacilityHoustonUSA
  3. 3.Department of Biostatistics and Applied MathematicsUniversity of Texas M. D. Anderson Cancer Center Genetically Engineered Mouse FacilityHoustonUSA

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