Abstract
Medullary thyroid carcinoma contributes to about 3–4% of thyroid cancers and affects C cells rather than follicular cells. Thyroid C cell differentiation from human pluripotent stem cells has not been reported. We report the stepwise differentiation of human embryonic stem cells into thyroid C cell-like cells through definitive endoderm and anterior foregut endoderm and ultimobranchial body-like intermediates in monolayer and 3D Matrigel culture conditions. The protocol involved sequential treatment with interferon/transferrin/selenium/pyruvate, foetal bovine serum, and activin A, then IGF-1 (Insulin-like growth factor 1), on the basis of embryonic thyroid developmental sequence. As well as expressing C cell lineage relative to follicular-lineage markers by qPCR (quantitative polymerase chain reaction) and immunolabelling, these cells by ELISA (enzyme-linked immunoassay) exhibited functional properties in vitro of calcitonin storage and release of calcitonin on calcium challenge. This method will contribute to developmental studies of the human thyroid gland and facilitate in vitro modelling of medullary thyroid carcinoma and provide a valuable platform for drug screening.
Highlights
1.1.IntroductionThyroid cancer a malignant tumour the endocrine system and the most commonThyroid cancer is is a malignant tumour ofof the endocrine system and the most common the endocrinecancers, cancers,accounting accountingforfor newly diagnosed cancers [1].Malignant ofofthe endocrine ofof newly diagnosed cancersMalignant thyroidcancers cancerscan canbe be categorised categorised as origin (papillary, folthyroid as differentiated differentiatedofoffollicular follicularcell cell origin, cell),less lessdifferentiated differentiatedofofCCcell cellorigin origin(medullaryand mostly hereditary [2], and undifferentiated and aggressive [3].[3]
Recent research has thrown light on the embryonic origin, development, and differentiation of mouse thyroid C cells in vivo, proving that C cells originate from foregut endodermal cells via the ubb [34]
We modified a previous published protocol on differentiation of definitive endoderm (DE) [39,40] by adding ITS-A to the media to support the growth of the cells in the presence of low FBS [26] (Figure 2a)
Summary
1.1.IntroductionThyroid cancer a malignant tumour the endocrine system and the most commonThyroid cancer is is a malignant tumour ofof the endocrine system and the most common the endocrinecancers, cancers,accounting accountingforfor newly diagnosed cancers [1].Malignant ofofthe endocrine ofof newly diagnosed cancersMalignant thyroidcancers cancerscan canbe be categorised categorised as origin (papillary, folthyroid as differentiated differentiatedofoffollicular follicularcell cell origin (papillary, licular, and rthlecell), cell),less lessdifferentiated differentiatedofofCCcell cellorigin origin(medullary (medullarythyroid thyroidcarcinoma carcinoma follicular, andH Hṻrthle oror MTC)and mostly hereditary [2], and undifferentiated and aggressive (anaplastic) [3].[3]. Thyroid cancer a malignant tumour the endocrine system and the most common. Thyroid cancer is is a malignant tumour ofof the endocrine system and the most common the endocrinecancers, cancers,accounting accountingforfor newly diagnosed cancers [1]. Malignant ofofthe endocrine ofof newly diagnosed cancers. Malignant thyroidcancers cancerscan canbe be categorised categorised as origin (papillary, folthyroid as differentiated differentiatedofoffollicular follicularcell cell origin (papillary, licular, and rthlecell), cell),less lessdifferentiated differentiatedofofCCcell cellorigin origin(medullary (medullarythyroid thyroidcarcinoma carcinoma follicular, andH Hṻrthle oror MTC). Mostly hereditary [2], and undifferentiated and aggressive (anaplastic) [3].[3].
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