Conn P M, Tsuruhara T, Dufau M, Catt K J
Endocrinology. 1977 Aug;101(2):639-42. doi: 10.1210/endo-101-2-639.
Purification of specific endocrine cells from mixed populations after dispersion of target tissues is important for detailed analysis of mechanisms of hormone action. A simple method for rapid isolation of endocrine cells with retention of biological integrity, has been developed by centrifugation in density gradients formed with Metrizamide. By this procedure, highly purified Leydig cells retaining morphological and biochemical characteristics were obtained. Such preparations bound 20, 300+/-3, 100 molecules of hCG per cell with affinity of 1.1+/-0.25 X 10(10) M-1. During incubation with hCG, cyclic AMP and testosterone responses of purified Leydig cells were considerably increased, and hCG concentrations as low as 0.2 pM caused activation of cAMP-dependent protein kinase.
在靶组织分散后从混合细胞群体中纯化特定内分泌细胞，对于详细分析激素作用机制至关重要。通过在由甲泛葡胺形成的密度梯度中离心，已开发出一种简单方法，可快速分离内分泌细胞并保持其生物学完整性。通过此程序，获得了保留形态和生化特征的高度纯化的睾丸间质细胞。此类制剂每个细胞结合20,300±3,100个hCG分子，亲和力为1.1±0.25×10¹⁰ M⁻¹。在与hCG孵育期间，纯化的睾丸间质细胞的环磷酸腺苷（cAMP）和睾酮反应显著增加，低至0.2 pM的hCG浓度即可激活cAMP依赖性蛋白激酶。
