The primary function of the thyroid gland is to metabolize iodide by synthesizing thyroid hormones, which are critical regulators of growth, development and metabolism in almost all tissues. So far, research on thyroid morphogenesis has been missing an efficient stem-cell model system that allows for the in vitro recapitulation of the molecular and morphogenic events regulating thyroid follicular-cell differentiation and subsequent assembly into functional thyroid follicles. Here we report that a transient overexpression of the transcription factors NKX2-1 and PAX8 is sufficient to direct mouse embryonic stem-cell differentiation into thyroid follicular cells that organize into three-dimensional follicular structures when treated with thyrotropin. These in vitro-derived follicles showed appreciable iodide organification activity. Importantly, when grafted in vivo into athyroid mice, these follicles rescued thyroid hormone plasma levels and promoted subsequent symptomatic recovery. Thus, mouse embryonic stem cells can be induced to differentiate into thyroid follicular cells in vitro and generate functional thyroid tissue.

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We thank G. Vassart, C. Blanpain and P. Vanderhaeghen for discussions and comments and V. Janssens for technical help. X.-H.L., A.M.D. and S.R. are supported in part by grants DK15070 and DK91016 from the National Institutes of Health. This work was supported by the Belgian Fonds de la Recherche Scientifique Medicale (FRSM[2]3_4_557_08 and [3]3_4598_12), Action de Recherche Concertée de la Communauté Française de Belgique (ARC N°AUWB-08/13-ULB10), Fonds d’Encouragement à la Recherche and grants from the Belgian National Fund for Scientific Research (FNRS). F.A. and D.F.K. are FNRS and Fund for Research in the Industry and the Agriculture (FRIA) research fellows, R.O. is an FNRS Postdoctoral Researcher and S.C. is an FNRS Senior Research Associate.

Author information


  1. Institute of Interdisciplinary Research in Molecular Human Biology (IRIBHM), Université Libre de Bruxelles, 808 route de Lennik, 1070 Brussels, Belgium

    • Francesco Antonica
    • , Dominika Figini Kasprzyk
    • , Robert Opitz
    •  & Sabine Costagliola
  2. Lillehei Heart Institute and Department of Pediatrics, University of Minnesota, Minneapolis, 55455 Minnesota, USA

    • Michelina Iacovino
    •  & Michael Kyba
  3. Department of Medicine, The University of Chicago, Chicago, 606370 Illinois, USA

    • Xiao-Hui Liao
    • , Alexandra Mihaela Dumitrescu
    •  & Samuel Refetoff
  4. Departments of Pediatrics & Genetics, The University of Chicago, Chicago, 60637 Illinois, USA

    • Samuel Refetoff
  5. Department of Veterinary Medical Imaging & Small Animal Orthopaedics, Faculty of Veterinary Medicine, Ghent University, B-9820 Merelbeke, Belgium

    • Kathelijne Peremans
  6. FNRS, ERASME, Université Libre de Bruxelles, 808 Route de Lennik, 1070 Brussels, Belgium

    • Mario Manto


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F.A. and S.C. developed the project, designed the experiments and analysed the data. F.A. performed most of the in vitro experiments and in vivo studies. D.F.K. provided technical help for the in vitro differentiation and functional characterization of the cells. M.I and M.K. provided A2Lox-Cre embryonic stem cells. X.-H.L. analysed blood TSH levels. A.M.D. read the manuscript and made experimental suggestions. S.R. provided suggestions and advice on the experimental procedures. K.P. performed the whole-body scan. M.M. performed body-temperature measurements. F.A., R.O. and S.C. wrote the manuscript. All authors read and approved the final manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Sabine Costagliola.

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