Derivation and comparison of C57BL/6 embryonic stem cells to a widely used 129 embryonic stem cell line (original) (raw)

Abstract

Typically, embryonic stem (ES) cells derived from 129 mouse substrains are used to generate genetically altered mouse models. Resulting chimeric mice were then usually converted to a C57BL/6 background, which takes at least a year, even in the case of speed congenics. In recent years, embryonic stem cells have been derived from various mouse strains. However, 129 ES cells are still widely used partially due to poor germline transmission of ES cells derived from other strains. Availability of highly germline-competent C57BL/6 ES cells would enormously facilitate generation of genetically altered mice in a pure C57BL/6 genetic background by eliminating backcrossing time, and thus significantly reducing associated costs and efforts. Here, we describe establishment of a C57BL/6 ES cell line (LK1) and compare its efficacy to a widely used 129SvJ ES cell line (GSI-1) in generating germline chimeras. In contrast to earlier studies, our data shows that highly germline-competent C57BL/6 ES cell lines can be derived using a simple approach, and thus support broader use of C57BL/6 ES cell lines for genetically engineered mouse models.

Access this article

Log in via an institution

Subscribe and save

• Starting from 10 chapters or articles per month
• Access and download chapters and articles from more than 300k books and 2,500 journals
• Cancel anytime View plans

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

• Amano H, Itakura K, Maruyama M et al (2006) Identification and targeted disruption of the mouse gene encoding ESG1 (PH34/ECAT2/DPPA5). BMC Dev Biol 6:11
  Article PubMed Google Scholar
• Auerbach W, Dunmore JH, Fairchild-Huntress V et al (2000) Establishment and chimera analysis of 129/SvEv- and C57BL/6-derived mouse embryonic stem cell lines. Biotechniques 29:1024–1028, 1030, 1032
  Google Scholar
• Brook FA, Gardner RL (1997) The origin and efficient derivation of embryonic stem cells in the mouse. Proc Natl Acad Sci USA 94:5709–5712
  Article PubMed CAS Google Scholar
• Chang JC, Ye L, Kan YW (2006) Correction of the sickle cell mutation in embryonic stem cells. Proc Natl Acad Sci USA 103:1036–1040
  Article PubMed CAS Google Scholar
• Cheng J, Dutra A, Takesono A et al (2004) Improved generation of C57BL/6J mouse embryonic stem cells in a defined serum-free media. Genesis 39:100–104
  Article PubMed Google Scholar
• Crawley JN, Belknap JK, Collins A et al (1997) Behavioral phenotypes of inbred mouse strains: implications and recommendations for molecular studies. Psychopharmacology (Berl) 132:107–124
  Article CAS Google Scholar
• Doetschman T, Gregg RG, Maeda N et al (1987) Targetted correction of a mutant HPRT gene in mouse embryonic stem cells. Nature 330:576–578
  Article PubMed CAS Google Scholar
• Evans MJ, Kaufman MH (1981) Establishment in culture of pluripotential cells from mouse embryos. Nature 292:154–156
  Article PubMed CAS Google Scholar
• Fedorov LM, Haegel-Kronenberger H, Hirchenhain J (1997) A comparison of the germline potential of differently aged ES cell lines and their transfected descendants. Transgenic Res 6:223–231
  Article PubMed CAS Google Scholar
• Hogan B, Costantini F, Lacy E (1994) Manipulating the mouse embryo: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, New York
  Google Scholar
• Kontgen F, Suss G, Stewart C et al (1993) Targeted disruption of the MHC class II Aa gene in C57BL/6 mice. Int Immunol 5:957–964
  Article PubMed CAS Google Scholar
• Ledermann B, Burki K (1991) Establishment of a germ-line competent C57BL/6 embryonic stem cell line. Exp Cell Res 197:254–258
  Article PubMed CAS Google Scholar
• Lemckert FA, Sedgwick JD, Korner H (1997) Gene targeting in C57BL/6 ES cells. Successful germ line transmission using recipient BALB/c blastocysts developmentally matured in vitro. Nucleic Acids Res 25:917–918
  Article PubMed CAS Google Scholar
• Linder CC (2001) The influence of genetic background on spontaneous and genetically engineered mouse models of complex diseases. Lab Anim (NY) 30:34–39
  CAS Google Scholar
• Liu P, Zhang H, McLellan A et al (1998) Embryonic lethality and tumorigenesis caused by segmental aneuploidy on mouse chromosome 11. Genetics 150:1155–1168
  PubMed CAS Google Scholar
• Martin GR (1981) Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc Natl Acad Sci USA 78:7634–7638
  Article PubMed CAS Google Scholar
• McVicar DW, Winkler-Pickett R, Taylor LS et al (2002) Aberrant DAP12 signaling in the 129 strain of mice: implications for the analysis of gene-targeted mice. J Immunol 169:1721–1728
  PubMed CAS Google Scholar
• Nagy A, Rossant J, Nagy R et al (1993) Derivation of completely cell culture-derived mice from early-passage embryonic stem cells. Proc Natl Acad Sci USA 90:8424–8428
  Article PubMed CAS Google Scholar
• Schuster-Gossler K, Lee AW, Lerner CP et al (2001) Use of coisogenic host blastocysts for efficient establishment of germline chimeras with C57BL/6J ES cell lines. Biotechniques 31:1022–1024, 1026
  Google Scholar
• Seong E, Saunders TL, Stewart CL et al (2004) To knockout in 129 or in C57BL/6: that is the question. Trends Genet 20:59–62
  Article PubMed CAS Google Scholar
• Shimizukawa R, Sakata A, Hirose M et al (2005) Establishment of a new embryonic stem cell line derived from C57BL/6 mouse expressing EGFP ubiquitously. Genesis 42:47–52
  Article PubMed CAS Google Scholar
• te Riele H, Maandag ER, Berns A (1992) Highly efficient gene targeting in embryonic stem cells through homologous recombination with isogenic DNA constructs. Proc Natl Acad Sci USA 89:5128–5132
  Article Google Scholar
• Thomas KR, Capecchi MR (1987) Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell 51:503–512
  Article PubMed CAS Google Scholar
• Tompers DM, Labosky PA (2004) Electroporation of murine embryonic stem cells: a step-by-step guide. Stem Cells 22:243–249
  Article PubMed Google Scholar
• Ware CB, Siverts LA, Nelson AM et al (2003) Utility of a C57BL/6 ES line versus 129 ES lines for targeted mutations in mice. Transgenic Res 12:743–746
  Article PubMed CAS Google Scholar
• Waterston RH, Lindblad-Toh K, Birney E et al (2002) Initial sequencing and comparative analysis of the mouse genome. Nature 420:520–562
  Article PubMed CAS Google Scholar
• Yang Y, Seed B (2003) Site-specific gene targeting in mouse embryonic stem cells with intact bacterial artificial chromosomes. Nat Biotechnol 21:447–451
  Article PubMed CAS Google Scholar
• Zhou L, Rowley DL, Mi QS et al (2001) Murine inter-strain polymorphisms alter gene targeting frequencies at the mu opioid receptor locus in embryonic stem cells. Mamm Genome 12:772–778
  Article PubMed CAS Google Scholar

Download references

Acknowledgements

We thank the Medical College of Georgia for funding and Dr. Pandelakis Koni for careful reading.

Author information

Author notes

1. Levent Keskintepe
   Present address: Sher Institute for Reproductive Medicine, Las Vegas, NV, USA

Authors and Affiliations

1. Mouse ES Cell and Transgenic Core Facility, Institute of Molecular Medicine and Genetics, Medical College of Georgia, 1120 15th Street, CB-2803, Augusta, GA, USA
   Levent Keskintepe, Karen Norris, Gabriela Pacholczyk, Suzanne Morris Dederscheck & Ali Eroglu
2. Department of Pathology, Medical College of Georgia, Augusta, GA, USA
   Karen Norris
3. Department of Medicine, Medical College of Georgia, Augusta, GA, USA
   Ali Eroglu

Authors

1. Levent Keskintepe
2. Karen Norris
3. Gabriela Pacholczyk
4. Suzanne Morris Dederscheck
5. Ali Eroglu

Corresponding author

Correspondence toAli Eroglu.

Electronic supplementary material

Rights and permissions

About this article

Cite this article

Keskintepe, L., Norris, K., Pacholczyk, G. et al. Derivation and comparison of C57BL/6 embryonic stem cells to a widely used 129 embryonic stem cell line.Transgenic Res 16, 751–758 (2007). https://doi.org/10.1007/s11248-007-9125-8

Download citation

• Received: 29 March 2007
• Accepted: 31 July 2007
• Published: 16 August 2007
• Issue date: December 2007
• DOI: https://doi.org/10.1007/s11248-007-9125-8

Keywords