本综述总结了离体研究肾脏嵌入细胞 (IC) 功能的方法。虽然 IC 功能障碍对于酸碱稳态很重要，但直到其变得严重时，临床上才会发现它。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。虽然体外肾小管灌注是研究 IC 的经典离体技术，但在这里我们专注于原代细胞培养物、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，这些信号通路可快速响应细胞外 pH、CO2 和碳酸氢盐 (HCO3-) 的变化。然而，这些 IC 工作方法也可能具有挑战性，因为重演 IC 的细胞系在培养物中不易增殖。此外，培养中的“纯”IC群体不一定复制其集合管（CD）环境，其中IC被更丰富的主细胞（PC）包围。令人欣慰的是，在离体 IC 系统信号传导中获得的许多发现在体内得到了很大程度上的证实。其中一些新发现的信号通路表明 IC 对于调节 NaCl 重吸收非常重要，从而为靶向抗高血压治疗开辟了新领域。此外，最近对肾上皮细胞的单细胞表征研究揭示了 IC 的双重发育起源，以及具有某些 IC 特征的新型 CD 细胞类型的存在。这些令人兴奋的发现为 IC 离体研究提供了新的机会，并且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 IC are surrounded by the more abundant principal cells (PC). 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 IC are important for regulating NaCl reabsorption, thus suggesting new frontiers to target anti-hypertensive 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.