title: Humanized medium (h7H) allows long-term primary follicular thyroid cultures from human normal thyroid, benign neoplasm, and cancer.
abstract: BACKGROUND
  Mechanisms of thyroid physiology and cancer are principally studied in follicular cell lines. However, human thyroid cancer lines were found to be heavily contaminated by other sources, and only one supposedly normal-thyroid cell line, immortalized with SV40 antigen, is available. In primary culture, human follicular cultures lose their phenotype after passage. We hypothesized that the loss of the thyroid phenotype could be related to culture conditions in which human cells are grown in medium optimized for rodent culture, including hormones with marked differences in its affinity for the relevant rodent/human receptor.
  
  
  OBJECTIVE
  The objective of the study was to define conditions that allow the proliferation of primary human follicular thyrocytes for many passages without losing phenotype.
  
  
  METHODS
  Concentrations of hormones, transferrin, iodine, oligoelements, antioxidants, metabolites, and ethanol were adjusted within normal homeostatic human serum ranges. Single cultures were identified by short tandem repeats. Human-rodent interspecies contamination was assessed.
  
  
  RESULTS
  We defined an humanized 7 homeostatic additives medium enabling growth of human thyroid cultures for more than 20 passages maintaining thyrocyte phenotype. Thyrocytes proliferated and were grouped as follicle-like structures; expressed Na+/I- symporter, pendrin, cytokeratins, thyroglobulin, and thyroperoxidase showed iodine-uptake and secreted thyroglobulin and free T3. Using these conditions, we generated a bank of thyroid tumors in culture from normal thyroids, Grave's hyperplasias, benign neoplasms (goiter, adenomas), and carcinomas.
  
  
  CONCLUSIONS
  Using appropriate culture conditions is essential for phenotype maintenance in human thyrocytes. The bank of thyroid tumors in culture generated under humanized humanized 7 homeostatic additives culture conditions will provide a much-needed tool to compare similarly growing cells from normal vs pathological origins and thus to elucidate the molecular basis of thyroid disease.
authors.affiliation: Centro de Investigaciones Medicas (CIMUS) e Instituto de Investigaciones Sanitarias and Department of Physiology, University of Santiago de Compostela and Complexo Hospitalario Universitario of Santiago de Compostela, 15782 Santiago de Compostela, Spain.
authors.forename: Susana B
authors.initials: SB
authors.lastname: Bravo
authors.forename: Maria E R
authors.initials: ME
authors.lastname: Garcia-Rendueles
authors.forename: Angela R
authors.initials: AR
authors.lastname: Garcia-Rendueles
authors.forename: Joana S
authors.initials: JS
authors.lastname: Rodrigues
authors.forename: Sihara
authors.initials: S
authors.lastname: Perez-Romero
authors.forename: Montserrat
authors.initials: M
authors.lastname: Garcia-Lavandeira
authors.forename: Maria
authors.initials: M
authors.lastname: Suarez-Fariña
authors.forename: Francisco
authors.initials: F
authors.lastname: Barreiro
authors.forename: Barbara
authors.initials: B
authors.lastname: Czarnocka
authors.forename: Ana
authors.initials: A
authors.lastname: Senra
authors.forename: Maria V
authors.initials: MV
authors.lastname: Lareu
authors.forename: Javier
authors.initials: J
authors.lastname: Rodriguez-Garcia
authors.forename: Jose
authors.initials: J
authors.lastname: Cameselle-Teijeiro
authors.forename: Clara V
authors.initials: CV
authors.lastname: Alvarez
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003470:Culture Media; D006801:Humans; D010641:Phenotype; D051381:Rats; D013954:Thyroglobulin / Q000378:metabolism; D013961:Thyroid Gland / Q000166:cytology*; D013964:Thyroid Neoplasms / Q000473:pathology*; D014284:Triiodothyronine / Q000378:metabolism
chemical_list: D003470:Culture Media; D014284:Triiodothyronine; D013954:Thyroglobulin