Humanized medium (h7H) allows long-term primary follicular thyroid cultures from human normal thyroid, benign neoplasm, and cancer
Bravo López, Susana Belén; Rodríguez García-Rendueles, María E; Rodríguez García-Rendueles, Ángela; Sousa Rodrigues, Joana; Pérez Romero, Sihara; García Lavandeira, Montserrat; Suárez Fariña, María; Barreiro Morandeira, Francisco; Czarnocka, B.; Senra ?, Ana; Lareu Huidobro, María Victoria; Rodríguez García, Javier; Cameselle Teijeiro, Jose Manuel; Álvarez Villamarín, Clara
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JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM
Type of content
MeSHAnimals | Cell Culture Techniques | Cell Line, Tumor | Cell Proliferation | Culture Media | Humans | Phenotype | Rats | Thyroglobulin | Thyroid Gland | Thyroid Neoplasms | Triiodothyronine
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.|The objective of the study was to define conditions that allow the proliferation of primary human follicular thyrocytes for many passages without losing phenotype.|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.|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.|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.