Ambesi-Impiombato F S, Parks L A, Coon H G
Proc Natl Acad Sci U S A. 1980 Jun;77(6):3455-9. doi: 10.1073/pnas.77.6.3455.
Primary cultures of rat thyroid cells were made in medium supplemented with 0.1--0.5% calf serum and containing six hormones or growth factors: insulin, thyrotropin, transferrin, hydrocortisone, somatostatin, and glycyl-L-histidyl-L-lysine acetate. The FRTL strain was purified by successive colonial isolations and was found to maintain highly differentiated features (secretion into the culture medium of physiological amounts of thyroglobulin and concentration of iodide by 100-fold). The FRTL strain has been observed for more than 3 years in continuous culture. It has maintained the same biochemical and morphological characteristics that typified the primary cultures of thyroid follicular cells immediately after their enzymatic release from the rat thyroid. Thyroid epithelial cells that were grown under more conventional cell culture conditions failed to retain these specialized characteristics. We show that maintenance in vitro of these specialized functions of rat thyroid follicular cells is dependent on low serum concentrations and supplementation with hormones in the primary cultures. Our observations indicate that this culture strategem may be aplicable to the general problem of maintenance of differentiated characteristics in cultures of other epithelial cells.
大鼠甲状腺细胞的原代培养是在添加了0.1 - 0.5%小牛血清且含有六种激素或生长因子的培养基中进行的,这六种激素或生长因子分别是:胰岛素、促甲状腺激素、转铁蛋白、氢化可的松、生长抑素和甘氨酰-L-组氨酰-L-赖氨酸乙酸盐。FRTL细胞系通过连续的克隆分离进行纯化,结果发现其能保持高度分化的特征(向培养基中分泌生理量的甲状腺球蛋白,并将碘化物浓缩100倍)。FRTL细胞系已在连续培养中观察了3年多。它保持了与刚从大鼠甲状腺经酶解分离出来的甲状腺滤泡细胞原代培养时相同的生化和形态特征。在更传统的细胞培养条件下生长的甲状腺上皮细胞未能保留这些特殊特征。我们表明,大鼠甲状腺滤泡细胞这些特殊功能在体外的维持依赖于原代培养中低血清浓度和激素补充。我们的观察表明,这种培养策略可能适用于维持其他上皮细胞培养中分化特征这一普遍问题。