DEPLETION OF L-3,5,3'-TRIIODOTHYRONINE AND L-THYROXINE IN EUTHYROID CALF SERUM FOR USE IN CELL-CULTURE STUDIES OF THE ACTION OF THYROID-HORMONE
DEPLETION OF L-3,5,3'-TRIIODOTHYRONINE AND L-THYROXINE IN EUTHYROID CALF SERUM FOR USE IN CELL-CULTURE STUDIES OF THE ACTION OF THYROID-HORMONE
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DOI：
10.1210/endo-105-1-80
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发表时间：
1979-01-01
期刊：
影响因子：
4.8
通讯作者：
CASANOVA, J
中科院分区：
文献类型：
--
作者：
SAMUELS, HH;STANLEY, F;CASANOVA, J
GH1细胞是大鼠垂体肿瘤细胞的克隆性株，能合成生长激素和催乳素。这些细胞对L-T3[三碘甲腺原氨酸]和L-T4[甲状腺激素]的生理浓度有反应，在细胞培养条件下，添加甲状腺切除的小牛血清以达到甲状腺激素耗竭状态。本文介绍了一种用阴离子交换树脂去除L-T3和L-T4正常甲状腺小牛血清的方法。该程序仅最小程度地改变了血清的低分子量阴离子成分，并且不改变总蛋白质含量或蛋白质的电泳图。用这种方法去除L-T_3和L-T_4的正常甲状腺小牛血清产生的血清，当用作介质补充时，其生物学反应与添加甲状腺切除的小牛血清所获得的生物反应相同。此外，如果甲状腺激素浓度恢复，树脂处理不会改变血清的促生长特性。在甲状腺切除不可行或需要对大量小动物进行外科手术的情况下，这种方法在准备用于细胞培养研究的甲状腺激素耗竭血清时应该是有用的。
GH1 cells are a clonal strain of rat pituitary tumor cells which synthesize GH [growth hormone] and PRL [prolactin]. These cells respond to physiological concentrations of L-T3 [triiodothyronine] and L-T4 [thyroxine] when cultured with medium supplemented with thyroidectomized calf serum to achieve a thyroid hormone-depleted state under cell culture conditions. A method to deplete euthyroid calf serum of L-T3 and L-T4 using an anion exhcange resin is described. The procedure only minimally alters the low MW anion components of the serum and does not change the total protein content or the electrophoretic pattern of proteins. Euthyroid calf serum depleted of L-T3 and L-T4 by this procedure yields serum which, when used as a medium supplement, results in biological responses identical to those obtained with media supplemented with thyroidectomized calf serum. In addition, resin treatment does not alter the growth-promoting properties of the serum if the thyroid hormone concentration is restored. This procedure should be useful in preparing thyroid hormone-depleted serum for cell culture studies in situations where thyroidectomy is not feasible or would require surgical procedures on a large number of small animals.