title: Differentiation of E14 mouse embryonic stem cells into thyrocytes in vitro.
abstract: 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.
authors.affiliation: Department of Nuclear Medicine, The Second Affiliated Hospital of Sun Yat-Sen University, GuangZhou, China. ningyij@163.net
authors.forename: Ningyi
authors.initials: N
authors.lastname: Jiang
authors.forename: Yingying
authors.initials: Y
authors.lastname: Hu
authors.forename: Xiongying
authors.initials: X
authors.lastname: Liu
authors.forename: Yanfeng
authors.initials: Y
authors.lastname: Wu
authors.forename: Hong
authors.initials: H
authors.lastname: Zhang
authors.forename: Guibing
authors.initials: G
authors.lastname: Chen
authors.forename: Jiugen
authors.initials: J
authors.lastname: Liang
authors.forename: Xianping
authors.initials: X
authors.lastname: Lu
authors.forename: Sheng
authors.initials: S
authors.lastname: Liu
mesh_terms: D000818:Animals; D002450:Cell Communication; D002454:Cell Differentiation / Q000187:drug effects*; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D005260:Female; D005347:Fibroblasts; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D000072636:PAX8 Transcription Factor; D051761:Paired Box Transcription Factors / Q000235:genetics / Q000378:metabolism; D011193:Potassium Iodide / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013961:Thyroid Gland / Q000166:cytology* / Q000648:ultrastructure; D000074482:Thyroid Nuclear Factor 1; D013972:Thyrotropin / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D007328:Insulin; C000614412:NKX2-1 protein, human; C000614414:Nkx2-1 protein, mouse; D009687:Nuclear Proteins; D000072636:PAX8 Transcription Factor; D051761:Paired Box Transcription Factors; C499496:Pax8 protein, mouse; D012333:RNA, Messenger; D000074482:Thyroid Nuclear Factor 1; D014157:Transcription Factors; D011193:Potassium Iodide; D013972:Thyrotropin