Huber G K, Davies T F
Department of Medicine, Mount Sinai School of Medicine, New York, New York 10029.
Endocrinology. 1990 Feb;126(2):869-75. doi: 10.1210/endo-126-2-869.
We have studied primary human fetal thyroid cell monolayers as an in vitro model for thyroid cell growth and function. Fetal thyroid cells grew slowly in a serum-free medium under basal conditions (i.e. without insulin and TSH), with a doubling time of 112 +/- 7 h (mean +/- SEM), indicating autonomous growth capacity. Addition of insulin (10 micrograms/ml) lead to increased growth, with a doubling time of 43.0 +/- 2.5 h, and TSH further reduced the doubling time in a dose-dependent manner, with the highest growth rate at 1-10 mU/ml (doubling time, 27 +/- 0.5 h). These growth rates were only observed when cells were subjected to a short culture time (12 h) before investigation, whereas after 96 h of culture the fetal thyroid cell growth rate was reduced by up to 50%. Addition of more than 5% serum completely inhibited the growth stimulation initiated by insulin and TSH. The accumulation of extracellular cAMP by the fetal cell monolayer was induced by TSH in a dose-dependent manner and reached a maximum effect at 10 mU/ml (4.8 +/- 0.6 pmol cAMP). Basal thyroglobulin (Tg) release was 26.9 +/- 0.5 ng/10(5) cells.day. Insulin decreased Tg release to 16.7 +/- 0.5 ng/10(5) cells,day, whereas TSH increased it up to 52.5 +/- 1.0 ng/10(5) cells.day. The T cell cytokine gamma-interferon, a product of the lymphocytic infiltrate in autoimmune thyroid disease, significantly reduced both insulin and TSH-stimulated cellular growth as well as accumulation of Tg. In conclusion, human fetal thyroid cell monolayers grew at high velocity under defined experimental conditions in vitro. These conditions included a low serum concentration, short culture time before investigation, and insulin/TSH supplementation. Furthermore, this human thyroid model confirms our earlier observations on the influence of cytokines in thyroid cell growth regulation.
我们研究了原代人胎儿甲状腺细胞单层，将其作为甲状腺细胞生长和功能的体外模型。在基础条件下（即无胰岛素和促甲状腺激素），胎儿甲状腺细胞在无血清培养基中生长缓慢，倍增时间为112±7小时（平均值±标准误），表明其具有自主生长能力。添加胰岛素（10微克/毫升）可促进生长，倍增时间为43.0±2.5小时，促甲状腺激素以剂量依赖方式进一步缩短倍增时间，在1 - 10毫国际单位/毫升时生长速率最高（倍增时间，27±0.5小时）。这些生长速率仅在细胞在研究前经过短时间培养（12小时）时观察到，而培养96小时后胎儿甲状腺细胞生长速率降低多达50%。添加超过5%的血清完全抑制了胰岛素和促甲状腺激素引发的生长刺激。胎儿细胞单层细胞外环磷酸腺苷（cAMP）的积累由促甲状腺激素以剂量依赖方式诱导，在10毫国际单位/毫升时达到最大效应（4.8±0.6皮摩尔cAMP）。基础甲状腺球蛋白（Tg）释放量为26.9±0.5纳克/10⁵细胞·天。胰岛素使Tg释放量降至16.7±0.5纳克/10⁵细胞·天，而促甲状腺激素使其增加至52.5±1.0纳克/10⁵细胞·天。T细胞细胞因子γ干扰素是自身免疫性甲状腺疾病中淋巴细胞浸润的产物，它显著降低了胰岛素和促甲状腺激素刺激的细胞生长以及Tg的积累。总之，人胎儿甲状腺细胞单层在体外特定实验条件下高速生长。这些条件包括低血清浓度、研究前短时间培养以及补充胰岛素/促甲状腺激素。此外，这个人类甲状腺模型证实了我们早期关于细胞因子对甲状腺细胞生长调节影响的观察结果。
