SpringerProtocols: Abstract: Culture of Murine Embryonic Stem Cells

11. Culture of Murine Embryonic Stem Cells

By: Ivana Barbaric², T. Neil Dear³

Abstract

Full Text

Download PDF (490K)

Appropriate culture of murine embryonic stem cells is critical to enabling introduced mutations to be passed through the germ line. ES cells must be carefully cultured to ensure that pluripotency is maintained. The feeder cells and foetal calf serum used to culture the cells can significantly influence germ line transmission potential. Additionally, ES cells can be karyotypically unstable, so determining the karyotype of ES cell lines will increase your confidence in the ability of the manipulated cells to contribute to the germ line in chimaeric mice.

Affiliation(s): (2) Department of Biomedical Science, University of Sheffield, Sheffield, UK
(3) Leeds Institute of Molecular Medicine, St. James’s University Hospital, Leeds, UK

Book Title: Transgenesis Techniques: Principles and Protocols

Series: Methods in Molecular Biology | Volume: 561 | Pub. Date: Jan-01-2009 | Page Range: 161-184 | DOI: 10.1007/978-1-60327-019-9_11

Subject: Genetics/Genomics

Key Words: Karyotype - LIF - MEF - Feeder cells - Freezing - Thawing

References

Download References

1.	Martin, G.R. (1981). Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc. Natl. Acad. Sci. U.S.A. 78, 7634–7638

2.	Evans, M.J. and Kaufman, M.H. (1981). Establishment in culture of pluripotential cells from mouse embryos. Nature 292, 154–156

3.	Williams, R.L., Hilton, D.J., Pease, S., Willson, T.A, Stewart, C.L., Gearing, D.P., Wagner, E.F., Metcalf, D., Nicola, N.A., and Gough, N.M. (1988). Myeloid leukaemia inhibitory factor maintains the developmental potential of embryonic stem cells. Nature 336, 684–687

4.	Smith, A.G., Heath, J.K., Donaldson, D.D., Wong, G.G., Moreau, J., Stahl, M., and Rogers, D. (1988). Inhibition of pluripotential embryonic stem cell differentiation by purified polypeptides. Nature 336, 688–690

5.	Solter, D. (2006). From teratocarcinomas to embryonic stem cells and beyond: a history of embryonic stem cell research. Nat. Rev. Genet. 7, 319–327

6.	Nagy, A. (2008). URL: www.mshri.on.ca/nagy/r1.htm retrieved on 28th February 2008

7.	Wellcome Trust Sanger Institute. (2008). URL: www.sanger.ac.uk/Software/archives/mouse-help.shtml retrieved on 28th February 2008

8.	Keskintepe, L., Norris, K., Pacholczyk, G., Dederscheck, S.M., and Eroglu, A. (2007). Derivation and comparison of C57BL/6 embryonic stem cells to a widely used 129 embryonic stem cell line. Transgenic Res. 16, 751–758

9.	Blake, M.S., Johnston, K.H., Russell-Jones, G.J., and Gotschlich, E.C. (1984). A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal. Biochem. 136, 175–179

10.	Ayral, A., Clarkson, S., Cheeseman, M., Wells, S., and Dear, T. (2006). A panel of optimized primers and positive control DNAs for PCR detection of common biological contaminants in mouse cell lines and tissue samples. Lab Anim. 35, 31–36

11.	Udy, G.B., Parkes, B.D., and Wells, D.N. (1997). ES cell cycle rates affect gene targeting frequencies. Exp. Cell Res.231, 296–301

12.	Balbach, S.T., Jauch, A., Böhm-Steuer, B., Cavaleri, F.M., Han, Y.M., and Boiani M. (2007). Chromosome stability differs in cloned mouse embryos and derivative ES cells. Dev. Biol. 308, 309–321

13.	Longo, L., Bygrave, A., Grosveld, F.G., and Pandolfi, P.P. (1997). The chromosome make-up of mouse embryonic stem cells is predictive of somatic and germ cell chimaerism. Transgenic Res. 6, 321–328

14.	Liu, X., Wu, H., Loring, J., Hormuzdi, S., Disteche, C.M., Bornstein, P., and Jaenisch R. (1997). Trisomy eight in ES cells is a common potential problem in gene targeting and interferes with germ line transmission. Dev. Dyn. 209, 85–91

15.	Hook, E.B. (1977). Exclusion of chromosomal mosaicism: Tables of 90%, 95%, and 99% confidence limits and comments on use. Am. J. Hum. Genet. 29, 94–97

16.	Nicklas, W., and Weiss. J. (2000). Survey of embryonic stem cells for murine infective agents. Comp. Med. 50, 410–411

17.	Kyuwa, S. (1997). Replication of murine coronaviruses in mouse embryonic stem cell lines in vitro. Exp. Anim. 46, 311–313

18.	Lindsey, J., Boorman, G., Collins, M., Hsu, C.-K., Van Hoosier., Jr.,G., and Wagner, J. (1991) Infectious Diseases in Mice and Rats. National Academy Press, Washington. DC. pp. 42–48

19.	Tucker, K.L., Wang, Y., Dausman, J., and Jaenisch, R. (1997). A transgenic mouse strain expressing four drug-selectable marker genes. Nucleic Acids Res.25, 3745–3746

20.	O’Gorman, S., Dagenais, N.A., Qian, M., and Marchuk, Y. (1997). Protamine-Cre recombinase transgenes efficiently recombine target sequences in the male germ line of mice, but not in embryonic stem cells. Proc. Natl. Acad. Sci. U.S.A. 94, 14602–14607

Comments (Loading...)

Post comment/View all comments