Embryonic Stem Cells/Induced Pluripotent Stem Cells

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Heparan Sulfation–Dependent Fibroblast Growth Factor Signaling Maintains Embryonic Stem Cells Primed for Differentiation in a Heterogeneous State†‡§

  1. Fredrik Lanner1,2,¶,*,
  2. Kian Leong Lee3,
  3. Marcus Sohl1,
  4. Katarina Holmborn4,
  5. Henry Yang5,
  6. Johannes Wilbertz1,
  7. Lorenz Poellinger1,3,
  8. Janet Rossant2,6,
  9. Filip Farnebo1

Article first published online: 23 NOV 2009

DOI: 10.1002/stem.265

Issue

STEM CELLS

STEM CELLS

Volume 28, Issue 2, pages 191–200, February 2010

Additional Information

How to Cite

Lanner, F., Lee, K. L., Sohl, M., Holmborn, K., Yang, H., Wilbertz, J., Poellinger, L., Rossant, J. and Farnebo, F. (2010), Heparan Sulfation–Dependent Fibroblast Growth Factor Signaling Maintains Embryonic Stem Cells Primed for Differentiation in a Heterogeneous State. STEM CELLS, 28: 191–200. doi: 10.1002/stem.265

Author Information

  1. 1

    Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden

  2. 2

    Program in Developmental and Stem Cell Biology, Research Institute, The Hospital for Sick Children, Toronto, Ontario, Canada

  3. 3

    Cancer Science Institute of Singapore, National University of Singapore, Singapore

  4. 4

    Department of Medical Biochemistry and Microbiology, Biomedical Centre, Uppsala University; Uppsala, Sweden

  5. 5

    Singapore Immunology Network, Biopolis, Agency for Science, Technology and Research, Singapore

  6. 6

    Department of Obstetrics and Gynecology, University of Toronto, Toronto, Ontario, Canada

  1. Telephone: 416-553-1308; Fax: 416-813-5252

*Program in Developmental and Stem Cell Biology, Research Institute, The Hospital for Sick Children, Toronto, Ontario, M5G 1X8, Canada

  1. †

    Author contributions: F.L.: Conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing, final approval of manuscript; K.L.L.: Collection and assembly of data, data analysis and interpretation; M.S.: Collection and assembly of data; K.H.: Provision of study material; H.Y.: Collection and assembly of data; J.W.: Data analysis and interpretation; L.P.: Provision of study material; J.R.: Conception and design, data analysis and interpretation, financial support, manuscript writing; F.F.: Conception and design, financial support, data analysis and interpretation, manuscript writing, final approval of manuscript.

  2. ‡

    Disclosure of potential conflicts of interest is found at the end of this article.

  3. §

    First published online in STEM CELLSEXPRESS January 28, 2010.

Publication History

  1. Issue published online: 16 FEB 2010
  2. Article first published online: 23 NOV 2009
  3. Accepted manuscript online: 23 NOV 2009 12:00AM EST
  4. Manuscript Accepted: 11 NOV 2009
  5. Manuscript Received: 27 AUG 2009

Funded by

  • Swedish Medical Research Council
  • Swedish Cancer Foundation
  • Swedish Heart Lung Foundation
  • Swedish Foundation for Strategic Research
  • Canadian Institutes for Health Research
  • Terry Fox Foundation

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Keywords:

  • Differentiation;
  • Embryonic stem cells;
  • Heparin;
  • Self-renewal

Abstract

Embryonic stem (ES) cells continuously decide whether to maintain pluripotency or differentiate. While exogenous leukemia inhibitory factor and BMP4 perpetuate a pluripotent state, less is known about the factors initiating differentiation. We show that heparan sulfate (HS) proteoglycans are critical coreceptors for signals inducing ES cell differentiation. Genetic targeting of NDST1 and NDST2, two enzymes required for N-sulfation of proteoglycans, blocked differentiation. This phenotype was rescued by HS presented in trans or by soluble heparin. NaClOmath image, which reduces sulfation of proteoglycans, potently blocked differentiation of wild-type cells. Mechanistically, N-sulfation was identified to be critical for functional autocrine fibroblast growth factor 4 (FGF4) signaling. Microarray analysis identified the pluripotency maintaining transcription factors Nanog, KLF2/4/8, Tbx3, and Tcf3 to be negatively regulated, whereas markers of differentiation such as Gbx2, Dnmt3b, FGF5, and Brachyury were induced by sulfation-dependent FGF receptor (FGFR) signaling. We show that several of these genes are heterogeneously expressed in ES cells, and that targeting of heparan sulfation or FGFR-signaling facilitated a homogenous Nanog/KLF4/Tbx3 positive ES cell state. This finding suggests that the recently discovered heterogeneous state of ES cells is regulated by HS-dependent FGFR signaling. Similarly, culturing blastocysts with NaClOmath image eliminated GATA6-positive primitive endoderm progenitors generating a homogenous Nanog-positive inner cell mass. Functionally, reduction of sulfation robustly improved de novo ES cell derivation efficiency. We conclude that N-sulfated HS is required for FGF4 signaling to maintain ES cells primed for differentiation in a heterogeneous state. Inhibiting this pathway facilitates a more naïve ground state. STEM CELLS 2010;28:191–200

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