Advanced in vitro kidney models are of great importance to the study of renal physiology and disease. Kidney tubuloids can be established from primary cells derived from adult kidney tissue or urine. Tubuloids are three-dimensional multicellular structures that recapitulate tubular function and have been used to study infectious, malignant, metabolic, and genetic diseases. For tubuloids to more closely represent the in vivo kidney, they can be integrated into an organ-on-a-chip system that has a more physiological tubular architecture and allows flow and interaction with vasculature or epithelial and mesenchymal cells from other organs. Here, we describe a detailed protocol for establishing tubuloid cultures from tissue and urine (1–3 weeks), as well as for generating and characterizing tubuloid cell–derived three-dimensional tubular structures in a perfused microfluidic multi-chip platform (7 d). The combination of the two systems yields a powerful in vitro tool that better recapitulates the complexity of the kidney tubule with donor-specific properties.

先进的体外肾脏模型对肾脏生理和疾病的研究具有重要意义。肾小管可以从源自成人肾组织或尿液的原代细胞中建立。Tubuloids 是概括肾小管功能的三维多细胞结构，已被用于研究传染性、恶性、代谢和遗传疾病。为了使肾小管更接近地代表体内肾脏，它们可以集成到具有更生理性的管状结构的器官芯片系统中，并允许与来自其他器官的脉管系统或上皮和间充质细胞流动和相互作用。在这里，我们描述了从组织和尿液（1-3 周）建立小管培养物的详细协议，以及在灌注微流体多芯片平台中生成和表征管状细胞衍生的三维管状结构 (7 d)。这两个系统的结合产生了一种强大的体外工具，可以更好地概括肾小管的复杂性和供体特异性。