Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Comparative Study
. 2008 Apr;58(2):145-50.

Comparison of tetraploid blastocyst microinjection of outbred Crl:CD1(ICR), hybrid B6D2F1/Tac, and inbred C57BL/6NTac embryos for generation of mice derived from embryonic stem cells

Sharron M Kirchain  1 , Alison M Hayward, John M Mkandawire, Peimin Qi, Aurora A Burds
Affiliations
Comparative Study

Comparison of tetraploid blastocyst microinjection of outbred Crl:CD1(ICR), hybrid B6D2F1/Tac, and inbred C57BL/6NTac embryos for generation of mice derived from embryonic stem cells

Sharron M Kirchain et al. Comp Med. 2008 Apr.

Abstract

Embryo electrofusion and tetraploid blastocyst microinjection is a modification of the traditional embryonic stem cell (ES cell)-based method to generate targeted mutant mice. Viability of tetraploid embryos is reportedly lower than with diploid embryos, with considerable interstrain variation. Here we assessed fetus and pup viability after ES cell microinjection of tetraploid blastocysts derived from outbred, hybrid, and inbred mice. Two-cell mouse embryos (C57BL/6NTac [B6], n = 788; B6D2F1/Tac [BDF1], n = 1871; Crl:CD1(ICR) [CD1], n = 1308) were electrofused; most resultant tetraploid blastocysts were injected with ES cells and surgically transferred into pseudopregnant recipient mice. Reproductive tracts were examined at midgestation for embryologic studies using B6 and BDF1 blastocysts; implantation sites and viable fetuses were counted. Pregnancies were carried to term for studies of targeted mutant mice using BDF1 and CD1 blastocysts, and pup yield was evaluated. Electrofusion rates of 2-cell embryos did not differ among B6, BDF1, and CD1 mice (overall mean, 92.8% +/- 5.4%). For embryologic studies, 244 B6 blastocysts were surgically transferred and 1 fetus was viable (0.41%), compared with 644 BDF1 blastocysts surgically transferred and 88 viable fetuses (13.7%). For targeted mutant mouse studies, 259 BDF1 blastocysts were surgically transferred yielding 10 pups (3.9%); 569 CD1 blastocysts yielded 44 pups (7.7%).

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
Implantation sites and fetal viability at E9.5–10.5 (B6 and BDF1 fetuses). For B6, 2 ES cell clones were used (uk and JW). For BDF1, 3 ES cell clones were used (JW, GFP, and B6). Of 244 B6 tetraploid blastocysts surgically transferred, 32 implantation sites were detected, and 1 fetus was viable. Of 294 BDF1 tetraploid blastocysts surgically transferred, 131 implantation sites were detected, and 29 fetuses were viable.
See this image and copyright information in PMC

References

    1. Aiba A, Inokuchi K, Ishida Y, Itohara S, Kobayashi K, Masu M, Mishina M, Miyakawa T, Mori H, Nakao K, Obata Y, Sakimura K, Shiroishi T, Wada K, Yagi T. 2007. Mouse liaison for integrative brain research. Neurosci Res 58:103–104 - PubMed
    1. Baharvand H, Matthaei KI. 2004. Culture condition difference for establishment of new embryonic stem cell lines from the C57Bl/6 and BALB/c mouse strains. In Vitro Cell Dev Biol Anim 40:76–81 - PubMed
    1. Bronson SK, Smithies O. 1994. Altering mice by homologous recombination using embryonic stem cells. J Biol Chem 269:27155–27159 - PubMed
    1. Cho M, Jang M, Lee EJ, Han JY, Lim JM. 2006. An alternative method of deriving embryonic stem cell-like clones by aggregation of diploid cells with tetraploid embryos. Fertil Steril 85Suppl 1:1103–1110 - PubMed
    1. Downing GJ, Battey JF., Jr 2004. Technical assessment of the first 20 years of research using mouse embryonic stem cell lines. Stem Cells 22:1168–1180 - PubMed

Publication types