Falke H E, Degenhart H J, Abeln G J, Visser H K, Croughs R J
Acta Endocrinol (Copenh). 1975 Jan;78(1):110-21. doi: 10.1530/acta.0.0780110.
A procedure for the continuous flow incubation of isolated adrenal cells is described. In this way the advantages of continuous flow incubations of adrenal tissue are combined with those of isolated adrenal cells. Suspensions of isolated adrenal cells were prepared by a modification of the collagenase method. A sigmoid dose-response curve was obtained when these cells were incubated with ACTH in batch incubations. Under these conditions (in the presence of 1 mU ACTH/ml) the corticosterone production rate remained constant during at least 240 min. This production rate was linearly related to the number of cells. Pre-incubation of the cells during 3 h resulted in an increased response to ACTH. In continuous flow incubations without ACTH the corticosterone production was negligible. With 100 mu U ACTH/ml corticosterone production increased sharply after a short lag period. A maximum was reached after 60-75 min followed by a slow decrease. Cells pre-incubated in the continuous flow apparatus had a slightly diminished ACTH response without loss of affinity to ACTH. The continuous flow incubation of isolated adrenal cells offers new possibilities for the dynamic study of steroid biosynthesis in vitro. The method may also be valuable to study processes in a wide variety of other tissues.
本文描述了一种分离肾上腺细胞的连续流动培养方法。通过这种方式，肾上腺组织连续流动培养的优点与分离肾上腺细胞的优点得以结合。分离肾上腺细胞悬液通过改良的胶原酶法制备。当这些细胞在批量培养中与促肾上腺皮质激素（ACTH）一起孵育时，得到了一条S形剂量反应曲线。在这些条件下（存在1 mU ACTH/ml），皮质酮产生率在至少240分钟内保持恒定。该产生率与细胞数量呈线性相关。细胞预孵育3小时导致对ACTH的反应增加。在无ACTH的连续流动培养中，皮质酮产生可忽略不计。加入100 μU ACTH/ml后，皮质酮产生在短暂延迟期后急剧增加。60 - 75分钟后达到最大值，随后缓慢下降。在连续流动装置中预孵育的细胞对ACTH的反应略有减弱，但对ACTH的亲和力未丧失。分离肾上腺细胞的连续流动培养为体外类固醇生物合成的动态研究提供了新的可能性。该方法对于研究多种其他组织中的过程可能也有价值。
