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An Ex vivo Culture System to Study Thyroid Development

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Thyroid follicle formation and thyroglobulin expression in multipotent endodermal stem cells.

Risheng Ma¹, Rauf Latif, Terry F Davies

¹Department of Medicine, Icahn School of Medicine at Mount Sinai, New York, New York, USA. risheng.ma@mssm.edu

Thyroid : Official Journal of the American Thyroid Association

|January 31, 2013

Summary

This summary is machine-generated.

Co-expressing thyroid transcription factors NKx2-1 and Pax8 in embryonic stem cells promotes thyroid follicular cell differentiation. This dual induction leads to the formation of functional thyroid cell structures in vitro.

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Area of Science:

Stem cell biology
Endocrinology
Developmental biology

Background:

Thyroid follicular cells (TFCs) are crucial for hormone production.
Thyroid transcription factors NKx2-1 and Pax8 are key regulators of TFC differentiation.
Embryonic stem cells (ES cells) offer a model for studying cell differentiation.

Purpose of the Study:

To investigate the impact of transcriptional induction of NKx2-1 and Pax8 on TFC differentiation from ES cells.
To assess the synergistic effects of co-expressed NKx2-1 and Pax8 on thyroid-specific gene activation.
To determine if dual transcription factor expression can commit ES cells to a TFC lineage.

Main Methods:

Murine ES cells were stably transfected with Pax8, NKx2-1, or both genes.
Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and protein expression analysis were used to assess gene induction.
Cells were differentiated into endodermal cells using activin A and TSH.

Main Results:

Co-expression of Pax8 and NKx2-1 significantly activated four key thyroid-specific genes: NIS, TSHR, Tg, and TPO.
Dual transfection in endodermally differentiated ES cells led to enhanced gene expression comparable to control thyroid cells.
Cells with dual transfection formed three-dimensional neofollicles in vitro and expressed Tg protein, unlike cells with single transcription factor expression.

Conclusions:

Co-expression of Pax8 and NKx2-1 in murine ES cells can induce thyroid-specific gene expression in endodermally differentiated cells.
This dual induction appears to commit ES cells to form three-dimensional neofollicular structures.
Findings support the role of these transcription factors in TFC differentiation from stem cells.