title: An Adult Mouse Thyroid Side Population Cell Line that Exhibits Enriched Epithelial-Mesenchymal Transition.
abstract: Studies of thyroid stem/progenitor cells have been hampered due to the small organ size and lack of tissue, which limits the yield of these cells. A continuous source that allows the study and characterization of thyroid stem/progenitor cells is desired to push the field forward.
  
  
  
  A cell line was established from Hoechst-resistant side population cells derived from mouse thyroid that were previously shown to contain stem/progenitor-like cells. Characterization of these cells were carried out by using in vitro two- and three-dimensional cultures and in vivo reconstitution of mice after orthotopic or intravenous injection, in conjunction with quantitative reverse transcription polymerase chain reaction, Western blotting, immunohisto(cyto)chemistry/immunofluorescence, and RNA seq analysis.
  
  
  
  These cells were named SPTL (side population cell-derived thyroid cell line). Under low serum culturing conditions, SPTL cells expressed the thyroid differentiation marker NKX2-1, a transcription factor critical for thyroid differentiation and function, while no expression of other thyroid differentiation marker genes were observed. SPTL cells formed follicle-like structures in Matrigel®cultures, which did not express thyroid differentiation marker genes. In mouse models of orthotopic and intravenous injection, the latter following partial thyroidectomy, a few SPTL cells were found in part of the follicles, most of which expressed NKX2-1. SPTL cells highly express genes involved in epithelial-mesenchymal transition, as demonstrated by RNA seq analysis, and exhibit a gene-expression pattern similar to anaplastic thyroid carcinoma.
  
  
  
  These results demonstrate that SPTL cells have the capacity to differentiate into thyroid to a limited degree. SPTL cells may provide an excellent tool to study stem cells, including cancer stem cells of the thyroid.
authors.affiliation: 1 Laboratory of Metabolism, National Cancer Institute, National Institutes of Health , Bethesda, Maryland.
authors.forename: Tsubasa
authors.initials: T
authors.lastname: Murata
authors.affiliation: 1 Laboratory of Metabolism, National Cancer Institute, National Institutes of Health , Bethesda, Maryland.
authors.forename: Manabu
authors.initials: M
authors.lastname: Iwadate
authors.affiliation: 1 Laboratory of Metabolism, National Cancer Institute, National Institutes of Health , Bethesda, Maryland.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Takizawa
authors.affiliation: 1 Laboratory of Metabolism, National Cancer Institute, National Institutes of Health , Bethesda, Maryland.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Miyakoshi
authors.affiliation: 1 Laboratory of Metabolism, National Cancer Institute, National Institutes of Health , Bethesda, Maryland.
authors.forename: Suguru
authors.initials: S
authors.lastname: Hayase
authors.affiliation: 2 DNA Sequencing and Genomics Core, National Heart, Lung, and Blood Institute , Bethesda, Maryland.
authors.forename: Wenjing
authors.initials: W
authors.lastname: Yang
authors.affiliation: 1 Laboratory of Metabolism, National Cancer Institute, National Institutes of Health , Bethesda, Maryland.
authors.forename: Yan
authors.initials: Y
authors.lastname: Cai
authors.affiliation: 1 Laboratory of Metabolism, National Cancer Institute, National Institutes of Health , Bethesda, Maryland.
authors.forename: Shigetoshi
authors.initials: S
authors.lastname: Yokoyama
authors.affiliation: 3 Electron Microscope Laboratory, Leidos Biomedical Research Inc. , Frederick National Laboratory for Cancer Research, Frederick, Maryland.
authors.forename: Kunio
authors.initials: K
authors.lastname: Nagashima
authors.affiliation: 2 DNA Sequencing and Genomics Core, National Heart, Lung, and Blood Institute , Bethesda, Maryland.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Wakabayashi
authors.affiliation: 4 Systems Biology Center, National Heart, Lung, and Blood Institute , Bethesda, Maryland.
authors.forename: Jun
authors.initials: J
authors.lastname: Zhu
authors.affiliation: 1 Laboratory of Metabolism, National Cancer Institute, National Institutes of Health , Bethesda, Maryland.
authors.forename: Shioko
authors.initials: S
authors.lastname: Kimura
mesh_terms: D000818:Animals; D015153:Blotting, Western; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D058750:Epithelial-Mesenchymal Transition; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D007150:Immunohistochemistry; D051379:Mice; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017423:Sequence Analysis, RNA; D058985:Side-Population Cells / Q000166:cytology* / Q000378:metabolism; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D065646:Thyroid Carcinoma, Anaplastic / Q000235:genetics; D013961:Thyroid Gland / Q000166:cytology*; D013964:Thyroid Neoplasms / Q000235:genetics; D000074482:Thyroid Nuclear Factor 1 / Q000378:metabolism; D013965:Thyroidectomy
chemical_list: C000614414:Nkx2-1 protein, mouse; D000074482:Thyroid Nuclear Factor 1
keywords: RNA seq analysis; epithelial-to-mesenchymal transition; thyroid cancer; thyroid side population; thyroid stem cells