Abstract
In order to study functions of normal human thyrocytes, we developed a protocol to obtain these cells in primary culture. Thyrocytes are obtained from normal tissue obtained at surgery for removal of thyroid neoplasms. Under sterile conditions, specimens are minced into small pieces, mono-dispersed cells are generated by digestion with collagenase type IV and the cells plated in tissue culture grade dishes in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS). After 24 h of incubation at 37 °C in a humidified 5% CO(2) incubator, the supernatant containing non-adherent cells is removed and the adherent cells are propagated in DMEM with 10% FBS, 100 IU/mL penicillin, and 10 μg/mL streptomycin. Cells proliferate with a doubling time of 72-94 h and retain functional characteristics for 9-12 doublings. We have used them successfully in studies to elucidate the signaling by thyrotropin (TSH) and insulin-like growth factor 1.