Guinea pig adrenocortical cells: in vitro characterization of separated zonal cell types.
Guinea pig adrenocortical cells: in vitro characterization of separated zonal cell types.
复制标题
豚鼠肾上腺皮质细胞：分离的带状细胞类型的体外表征。
DOI：
10.1083/jcb.94.2.241
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发表时间：
1982
期刊：
影响因子：
--
通讯作者：
Black,VH
中科院分区：
文献类型：
--
作者：
Miao,P;Black,VH
本文报道了一种快速、相对简单和可重复的技术，用于获得高达80-90%纯度的束状细胞和球状细胞群体，这些细胞可以在体外保持用于肾上腺皮质细胞功能的研究。使用平衡密度离心（570 g，30 min）在1-28%牛血清白蛋白/不含Ca++、Mg++的缓冲液梯度（wt/vol，增量为4%）上分离分离豚鼠肾上腺皮质细胞。分离后，在总分离物中获得的8 × 10（5）个活细胞/肾上腺中，回收率超过60%。80%的肾小球细胞位于梯度的下三带。78%的束状体细胞出现在前三条带中。在前两条带的细胞中，78-91%为束状细胞，而在后两条带的细胞中，92-95%为球状细胞。细胞保持了原位细胞的形态特征，并可在体外维持长达11 d。他们产生了各种各样的类固醇，皮质醇，皮质酮，醛固酮，11-β-羟基雄烯二酮，脱氧皮质醇，脱氧皮质酮，可的松，18-羟基皮质酮，以及一种暂时被鉴定为脱氢表雄酮的产物，他们以剂量反应的方式对ACTH产生反应，类固醇输出水平提高。肾小球带丰富的人口不同的束状体丰富的人口在其丰富的生产醛固酮和类固醇生产的模式。没有一种培养物对血管紧张素II（100 pg/ml）有反应，类固醇产生增加。
This paper reports a quick, relatively simple and reproducible technique for obtaining populations of zona fasciculata and zona glomerulosa cells up to 80-90% pure, which can be maintained in vitro for study of adrenocortical cell function. Isolated guinea pig adrenocortical cells were separated on a 1-28% bovine serum albumin/Ca++, Mg++-free buffer gradient (wt/vol at 4% increments) using equilibrium density centrifugation (570 g, 30 min). Over 60% of the 8 x 10(5) viable cells/adrenal obtained in the total isolate were recovered after separation. 80% of the zona glomerulosa cells were found in the lower three bands of the gradient. 78% of the zona fasciculata cells were found in the top three bands. Of the cells in the first two bands, 78-91% were zona fasciculata cells, whereas of the cells in the bottom two bands 92-95% were zona glomerulosa cells. The cells retained the morphological characteristics of cells in situ and could be maintained in vitro for periods up to 11 d. They produced a wide variety of steroids, cortisol, corticosterone, aldosterone, 11-beta-hydroxyandrostenedione, deoxycortisol, deoxycorticosterone, cortisone, 18-hydroxycorticosterone, and a product tentatively identified as dehydroepiandrosterone, and they responded to ACTH in a dose-responsive manner with enhanced levels of steroid output. Zona glomerulosa-enriched populations differed from zona fasciculata-enriched populations in their abundant production of aldosterone and in the pattern of steroid production. None of the cultures responded to angiotensin II (100 pg/ml) with increased steroid production.