Abstract
During the last decade three-dimensional (3D) cultures of pluripotent stem cells have been intensively used to understand morphogenesis and molecular signaling important for the embryonic development of many tissues. In addition, pluripotent stem cells have been shown to be a valid tool for the in vitro modeling of several congenital or chronic human diseases, opening new possibilities to study their physiopathology without using animal models. Even more interestingly, 3D culture has proved to be a powerful and versatile tool to successfully generate functional tissues ex vivo. Using similar approaches, we here describe a protocol for the generation of functional thyroid tissue using mouse embryonic stem cells and give all the details and references for its characterization and analysis both in vitro and in vivo. This model is a valid approach to study the expression and the function of genes involved in the correct morphogenesis of thyroid gland, to elucidate the mechanisms of production and secretion of thyroid hormones and to test anti-thyroid drugs.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Eiraku M, Takata N, Ishibashi H et al (2011) Self-organizing optic-cup morphogenesis in three-dimensional culture. Nature 472:51–56
Suga H, Kadoshima T, Minaguchi M et al (2011) Self-formation of functional adenohypophysis in three-dimensional culture. Nature 480:57–62
Nakano T, Ando S, Takata N et al (2012) Self-formation of optic cups and storable stratified neural retina from human ESCs. Cell Stem Cell 10:771–785
Antonica F, Kasprzyk DF, Opitz R et al (2012) Generation of functional thyroid from embryonic stem cells. Nature 491:66–71
Arufe MC, Lu M, Kubo A et al (2006) Directed differentiation of mouse embryonic stem cells into thyroid follicular cells. Endocrinology 147:3007–3015
Longmire TA, Ikonomou L, Hawkins F et al (2012) Efficient derivation of purified lung and thyroid progenitors from embryonic stem cells. Cell Stem Cell 10:398–411
Kyba M, Perlingeiro RC, Daley GQ (2002) HoxB4 confers definitive lymphoid-myeloid engraftment potential on embryonic stem cell and yolk sac hematopoietic progenitors. Cell 109:29–37
Ahfeldt T, Schinzel RT, Lee YK et al (2012) Programming human pluripotent stem cells into white and brown adipocytes. Nat Cell Biol 14:209–219
Kubo A, Stull R, Takeuchi M et al (2011) Pdx1 and Ngn3 overexpression enhances pancreatic differentiation of mouse ES cell-derived endoderm population. PLoS One 6:e24058
Kimura S, Hara Y, Pineau T et al (1996) The T/ebp null mouse: thyroid-specific enhancer-binding protein is essential for the organogenesis of the thyroid, lung, ventral forebrain, and pituitary. Genes Dev 10:60–69
Plachov D, Chowdhury K, Walther C et al (1990) Pax8, a murine paired box gene expressed in the developing excretory system and thyroid gland. Development 110:643–651
De Felice M, Di Lauro R (2004) Thyroid development and its disorders: genetics and molecular mechanisms. Endocr Rev 25:722–746
De Felice M, Di Lauro R (2011) Minireview: intrinsic and extrinsic factors in thyroid gland development: an update. Endocrinology 152:2948–2956
Kurmann AA, Serra M, Hawkins F et al (2015) Regeneration of thyroid function by transplantation of differentiated pluripotent stem cells. Cell Stem Cell 17:527–542
Iacovino M, Bosnakovski D, Fey H et al (2011) Inducible cassette exchange: a rapid and efficient system enabling conditional gene expression in embryonic stem and primary cells. Stem Cells 29:1580–1588
Hsiao EC, Nguyen TD, Ng JK et al (2011) Constitutive Gs activation using a single-construct tetracycline-inducible expression system in embryonic stem cells and mice. Stem Cell Res Ther 2:11
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Antonica, F., Kasprzyk, D.F., Schiavo, A.A., Romitti, M., Costagliola, S. (2017). Generation of Functional Thyroid Tissue Using 3D-Based Culture of Embryonic Stem Cells. In: Tsuji, T. (eds) Organ Regeneration. Methods in Molecular Biology, vol 1597. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6949-4_7
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6949-4_7
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6947-0
Online ISBN: 978-1-4939-6949-4
eBook Packages: Springer Protocols