本文综述了体外研究肾脏插入细胞(IC)功能的方法。虽然对酸碱稳态很重要,IC功能障碍通常在临床上无法识别,直到其变得严重。使用离体技术的优点是它们允许在受控环境中对IC功能进行差异评估。尽管体外肾小管灌注是研究IC的经典离体技术,这里我们专注于原代细胞培养,永生化细胞系,和离体肾切片。离体技术可用于评估IC信号通路,允许对pH的细胞外变化做出快速反应,CO2和碳酸氢盐(HCO3-)。然而,这些集成电路工作的方法也可能是具有挑战性的,因为概括IC的细胞系在培养中不容易增殖。此外,文化中的“纯”IC种群不一定复制其收集管(CD)环境,其中IC被更丰富的主细胞(PC)包围。令人放心的是,在离体IC系统信号传导中获得的许多发现已经在体内得到了大量证实。这些新发现的信号通路中的一些揭示了IC对于调节NaCl重吸收是重要的,因此提出了针对抗高血压治疗的新领域。此外,最近的肾上皮细胞的单细胞表征研究揭示了IC的双重发育起源,以及存在具有某些IC特征的新型CD细胞类型。这些令人兴奋的发现为IC离体研究提供了新的机会,并可能会重新发现该领域可用工具的重要性。
This review summarizes methods to
study kidney intercalated cell (IC) function ex vivo. While important for acid-base homeostasis, IC dysfunction is often not recognized clinically until it becomes severe. The advantage of using ex vivo techniques is that they allow for the differential evaluation of IC function in controlled environments. Although in vitro kidney tubular perfusion is a classical ex vivo technique to
study IC, here we concentrate on primary cell cultures, immortalized cell lines, and ex vivo kidney slices. Ex vivo techniques are useful in evaluating IC signaling pathways that allow rapid responses to extracellular changes in pH, CO2, and bicarbonate (HCO3-). However, these methods for IC work can also be challenging, as cell lines that recapitulate IC do not proliferate easily in culture. Moreover, a \"pure\" IC population in culture does not necessarily replicate its collecting duct (CD) environment, where ICs are surrounded by the more abundant principal cells (PCs). It is reassuring that many findings obtained in ex vivo IC systems signaling have been largely confirmed in vivo. Some of these newly identified signaling pathways reveal that ICs are important for regulating NaCl reabsorption, thus suggesting new frontiers to target antihypertensive treatments. Moreover, recent single-cell characterization studies of kidney epithelial cells revealed a dual developmental origin of IC, as well as the presence of novel CD cell types with certain IC characteristics. These exciting findings present new opportunities for the
study of IC ex vivo and will likely rediscover the importance of available tools in this field.NEW & NOTEWORTHY The
study of kidney intercalated cells has been limited by current cell culture and kidney tissue isolation techniques. This review is to be used as a reference to select ex vivo techniques to
study intercalated cells. We focused on the use of cell lines and kidney slices as potential useful models to study membrane transport proteins. We also review how novel collecting duct organoids may help better elucidate the role of these intriguing cells.