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Human Embryonic Stem Cell Protocols pp 267–283Cite as

Microarray Approach to Identify the Signaling Network Responsible for Self-Renewal of Human Embryonic Stem Cells

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Part of the book series: Methods In Molecular Biology ((MIMB,volume 331))

Abstract

This chapter introduces the representative method to culture human embryonic stem cells (hESCs) under the feeder and feeder-free conditions, the former of which is used to maintain or expand undifferentiated hESCs, and the latter can be used for preparation of pure hESCs RNA samples, or for screening factors influential on self-renewal of hESCs. Also described in this chapter: a protocol and tips for conducting gene chip analysis focusing on widely used Affymetrix microarrays. These techniques will provide an unprecedented scale of biological information to illuminate a key to decipher complex signaling networks controlling pluripotency.

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References

  1. Smith A. G. (2001) Embryo-derived stem cells: of mice and men. Annu. Rev. Cell. Dev. Biol. 17, 435–462.

    Article  PubMed  CAS  Google Scholar 

  2. Weissman I. L. (2000) Stem cells: units of development, units of regeneration, and units in evolution. Cell 100, 157–168.

    Article  PubMed  CAS  Google Scholar 

  3. Rossant J. (2001) Stem cells from the Mammalian blastocyst. Stem Cells 19, 477–482.

    Article  PubMed  CAS  Google Scholar 

  4. 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. USA 78, 7634–7638.

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  6. Hubner K., Fuhrmann G., Christenson L. K., et al. (2003) Derivation of oocytes from mouse embryonic stem cells. Science 300, 1251–1256.

    Article  PubMed  Google Scholar 

  7. Geijsen N., Horoschak M., Kim K., Gribnau J., Eggan K., and Daley G. Q. (2004) Derivation of embryonic germ cells and male gametes from embryonic stem cells. Nature 427, 148–154.

    Article  PubMed  CAS  Google Scholar 

  8. Kaufman M. H., Robertson E. J., Handyside A. H., and Evans M. J. (1983) Establishment of pluripotential cell lines from haploid mouse embryos. J. Embryol. Exp. Morphol. 73, 249–261.

    PubMed  CAS  Google Scholar 

  9. Thomson J. A., Itskovitz-Eldor J., Shapiro S. S., et al. (1998) Embryonic stem cell lines derived from human blastocysts. Science 282, 1145–1147.

    Article  PubMed  CAS  Google Scholar 

  10. Reubinoff B. E., Pera M. F., Fong C. Y., Trounson A., and Bongso A. (2000) Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro. Nat. Biotechnol. 18, 399–404.

    Article  PubMed  CAS  Google Scholar 

  11. Okamoto K., Okazawa H., Okuda A., Sakai M., Muramatsu M., and Hamada H. (1990) A novel octamer binding transcription factor is differentially expressed in mouse embryonic cells. Cell 60, 461–472.

    Article  PubMed  CAS  Google Scholar 

  12. Nichols J., Zevnik B., Anastassiadis K., et al. (1998) Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell 95, 379–391.

    Article  PubMed  CAS  Google Scholar 

  13. Scholer H. R., Ruppert S., Suzuki N., Chowdhury K., and Gruss P. (1990) New type of POU domain in germ line-specific protein Oct-4. Nature 344, 435–439.

    Article  PubMed  CAS  Google Scholar 

  14. Rogers M. B., Hosler B. A., and Gudas, L. J. (1991) Specific expression of a retinoic acid-regulated, zinc-finger gene, Rex-1, in preimplantation embryos, trophoblast and spermatocytes. Development 113, 815–824.

    PubMed  CAS  Google Scholar 

  15. Mitsui K., Tokuzawa Y., Itoh H., et al. (2003) he homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell 113, 631–642.

    Article  PubMed  CAS  Google Scholar 

  16. Chambers I., Colby D., Robertson M., et al. (2003) Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 113, 643–655.

    Article  PubMed  CAS  Google Scholar 

  17. Henderson J. K., Draper J. S., Baillie H. S., et al. (2002) Preimplantation human embryos and embryonic stem cells show comparable expression of stage-specific embryonic antigens. Stem Cells 20, 329–337.

    Article  PubMed  CAS  Google Scholar 

  18. Xu R. H., Chen X., Li D. S., et al. (2002) BMP4 initiates human embryonic stem cell differentiation to trophoblast. Nat. Biotechnol. 20, 1261–1264.

    Article  PubMed  CAS  Google Scholar 

  19. Niwa H., Miyazaki J., and Smith, A. G. (2000) Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells. Nat. Genet. 24, 372–376.

    Article  PubMed  CAS  Google Scholar 

  20. Ying Q. L., Nichols J., Chambers I., and Smith A. (2003) BMP induction of Id proteins suppresses differentiation and sustains embryonic stem cell self-renewal in collaboration with STAT3. Cell 115, 281–292.

    Article  PubMed  CAS  Google Scholar 

  21. Sato N., Sanjuan I. M., Heke M., Uchida M., Naef F., and Brivanlou A. H. (2003) Molecular signature of human embryonic stem cells and its comparison with the mouse. Dev. Biol. 260, 404–413.

    Article  PubMed  CAS  Google Scholar 

  22. Xu C., Inokuma M. S., Denham J., et al. (2001) Feeder-free growth of undifferentiated human embryonic stem cells. Nat. Biotechnol. 19, 971–974.

    Article  PubMed  CAS  Google Scholar 

  23. Draper J. S., Smith K., Gokhale P., et al. (2004) Recurrent gain of chromosomes 17q and 12 in cultured human embryonic stem cells. Nat. Biotechnol. 22, 53–54.

    Article  PubMed  CAS  Google Scholar 

  24. Baldi P. and Long A. D. (2001) A Bayesian framework for the analysis of microarray expression data: regularized t-test and statistical inferences of gene changes. Bioinformatics 17, 509–519.

    Article  PubMed  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. Laboratory of Molecular Embryology, The Rockefeller University, New York, NY

    Noboru Sato & Ali H. Brivanlou

Authors
  1. Noboru Sato
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  2. Ali H. Brivanlou
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Editor information

Editors and Affiliations

  1. Ottawa Health Research Institute, Ottawa, Ontario, Canada

    Kursad Turksen

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© 2006 Humana Press Inc.

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Sato, N., Brivanlou, A.H. (2006). Microarray Approach to Identify the Signaling Network Responsible for Self-Renewal of Human Embryonic Stem Cells. In: Turksen, K. (eds) Human Embryonic Stem Cell Protocols. Methods In Molecular Biology, vol 331. Humana Press. https://doi.org/10.1385/1-59745-046-4:267

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  • DOI: https://doi.org/10.1385/1-59745-046-4:267

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-497-5

  • Online ISBN: 978-1-59745-046-1

  • eBook Packages: Springer Protocols

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