Stem cells derived from Goiters in adults form spheres in response to intense growth stimulation and require thyrotropin for differentiation into thyrocytes

Objective: This study aimed to analyze under which conditions quiescent stem cells derived from human goiters can be propagated to outgrow and whether these cells have retained the capacity to differentiate into thyroid cells. Design: Stem cells were isolated by fluorescence-activated cell sorting as a side population by the Hoechst 33342 efflux technique. Growth pattern of stem cells and cocultures of stem cells with thyrocytes grown as monolayer and in Matrigel was investigated. Expression of stem cell markers, endodermal markers, and thyroidspecific markers was analyzed by RT-PCR. In stem cell-derived thyrocytes, embedded in collagen to form follicles, TSH-dependent (125)iodide uptake was measured. Results: Stem cells were isolated as a side population from a non-side population fraction that consisted of endodermal marker-positive cells and thyroid cells. Intense growth stimulation of stem cells in coculture with thyrocytes resulted in formation of nonadherent, three-dimensional spheres that consisted of highly proliferating stem cells with their characteristic expression profiles. In response to TSH and serum, sphere-derived progenitor cells differentiated into thyrocytes that expressed paired box gene 8, thyroglobulin, sodium iodide symporter, thyroid-stimulating hormone receptor, and thyroperoxidase mRNA and showed TSH-dependent 125iodide uptake. Conclusion: Quiescent stem cells derived from goiters can be propagated to form spheres that consist of highly proliferating stem cells that are able to differentiate TSH dependently into thyroid cells. Compared with thyrocytes, stem cells display a much higher proliferation rate on acute growth stimulation, which may suggest a putative role of the offspring of stem cells in the chronic growth factor-stimulated nodular transformation of the thyroid.