Differentiation of E14 mouse embryonic stem cells into thyrocytes in vitro.

BACKGROUND: If methods of differentiating stem cells into thyrocytes can be perfected, they may provide a ready source of normal thyrocytes for basic research and clinical application. We developed a novel culture method capable of differentiating mouse embryonic stem (ES) cells into thyroid follicular cells.
METHODS: E14 mouse ES cells were allowed to differentiate into embryoid bodies and then stimulated with thyroid-stimulating hormone, insulin, and potassium iodide. The resulting differentiated cells were observed for expression of thyrocyte-specific mRNA transcripts with reverse transcriptase (RT)-polymerase chain reaction. To definitively identify thyrocytes, we simultaneously observed the thyrocyte-specific proteins, thyroid transcription factor-1 and PAX-8, with dual-color immunofluorescent labeling. The cells were further characterized by electron microscopy.
RESULTS: The ES cells were successfully differentiated into thyrocytes. Differentiated cells expressed PAX-8, thyroid-stimulating hormone receptor, sodium/iodide symporter, thyroperoxidase, and thyroglobulin mRNAs, and coexpressed thyroid transcription factor-1 and PAX-8 proteins. The extent of differentiation was further explored by electron microscopy, which showed that differentiated cells had ultrastructural features similar to adult human thyrocytes, whereas the cells from unstimulated cultures were mostly disintegrated and lacked developed organelle structures.
CONCLUSIONS: These data show that E14 mouse ES cells can be differentiated into thyrocytes by culturing with thyroid-stimulating hormone, insulin, and potassium iodide. The development of reliable methods to produce thyroid cells from ES cells is important to future research in thyroid biology and medical applications.