Chen Yung-Chia, Huang Bu-Miin
Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University; Department of Anatomy, School of Medicine, Kaohsiung Medical University;
Department of Cell Biology and Anatomy, College of Medicine, National Cheng Kung University.
J Vis Exp. 2019 Mar 12(145). doi: 10.3791/59016.
The hormone which the adrenal cortex secretes is vital for animals against stress and diseases. The method described here is the procedure of primary cultured rat adrenal cells and related functional assays (immunofluorescence staining of lipid droplet surface protein, as well as corticosterone analysis). Unlike an in vivo model, the variation of interexperiments in adrenal monolayer cultures is less and the experimental condition is easy to control. Besides, the source of rats is also more stable than other animals, like bovine ones. There are also several human adrenal cell lines (NCI-H295, NCI-H295R, SW13, etc.) that can be used in adrenal studies. However, the steroid production of these lines will still be influenced by numerous factors, which include serum lot number, passage number, mutant/loss of distinct genes, etc. Except for lacking 17α-hydroxylase, the primary culture of rat adrenocortical cells is a better and more convenient technique for studying adrenal physiology. In summary, primary rat adrenal cultures could be a good in vitro platform for researchers to investigate the mechanisms of the reagent of interest in the adrenal gland system.
肾上腺皮质分泌的激素对动物抵抗压力和疾病至关重要。这里描述的方法是原代培养大鼠肾上腺细胞的程序以及相关功能测定(脂滴表面蛋白的免疫荧光染色以及皮质酮分析)。与体内模型不同,肾上腺单层培养实验间的差异较小,且实验条件易于控制。此外,大鼠的来源也比其他动物(如牛)更稳定。也有几种人肾上腺细胞系(NCI-H295、NCI-H295R、SW13等)可用于肾上腺研究。然而,这些细胞系的类固醇生成仍会受到众多因素的影响,包括血清批次号、传代次数、特定基因的突变/缺失等。除了缺乏17α-羟化酶外,大鼠肾上腺皮质细胞的原代培养是研究肾上腺生理学更好且更便捷的技术。总之,原代大鼠肾上腺培养可为研究人员提供一个良好的体外平台,用于研究肾上腺系统中感兴趣试剂的作用机制。