一种用于研究钠和水运输机制的新型人体远端微管芯片模型。

IF 3 Q1 UROLOGY & NEPHROLOGY
Kidney360 Pub Date : 2025-09-17 DOI:10.34067/KID.0000000992
Murillo D L Bernardi, Emre Dilmen, Dorota Kurek, Henriette L Lanz, Jos Joore, Joost G Hoenderop, Maarten B Rookmaaker, Marianne C Verhaar
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引用次数: 0

摘要

背景:远端肾元包括厚升肢、远曲小管、连接小管和收集管,在调节水电解质平衡中起重要作用。这些节段的损伤或功能障碍可导致严重的肾脏疾病和全身并发症。直接在体内研究这些区域受到肾脏结构复杂性的限制。最近,人肾小管细胞(称为小管)的三维培养和支持动态流动的微流控平台为体外肾脏功能建模提供了新的机会。方法:将原代人小管细胞与OrganoPlate®微流体培养系统结合,建立人远端微管细胞芯片模型。将小管注入到I型胶原基质附近形成三维管状结构,并暴露于交替流动中。分化为远端肾元表型的评估使用定量聚合酶链反应,免疫组织化学和屏障完整性分析。通过放射性标记的钠摄取、经上皮抵抗、葡聚糖扩散和圆顶形成来研究钠和水的功能性运输。药理研究使用了甲氧苄啶,一种上皮钠通道活性抑制剂。结果:与传统的二维培养相比,流动条件下的三维培养显示远端肾元段特异性标记物的表达增加。免疫染色证实形成了高度极化的上皮,关键转运蛋白定位于根尖和基底外侧。小管形成了一个密漏屏障,表现为葡聚糖渗透性降低和经皮细胞阻力升高。放射标记试验显示,活性钠转运由根尖上皮钠通道和基底外侧钠钾atp酶驱动。水的输送伴随着钠的移动,在上皮下形成穹窿。暴露于甲氧苄氨嘧啶减少钠摄取和圆顶形成,证实了模型的功能反应性。结论:这种远端小管芯片系统再现了人类远端肾元的关键结构和功能特征。它使研究盐和水在受控微流控下在原代人小管细胞中的运输成为可能。该模型为研究肾脏生理、病理和药理反应提供了一个生理学相关的可扩展平台,在药物发现和毒性测试中具有潜在的应用前景。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A Novel Human Distal Tubuloid-on-a-Chip Model for Investigating Sodium and Water Transport Mechanisms.

Background: The distal nephron, which includes the thick ascending limb, distal convoluted tubule, connecting tubule, and collecting duct, is essential for the regulation of water and electrolyte balance. Injury or dysfunction of these segments contributes to significant kidney disease and systemic complications. Studying these regions directly in vivo is limited by the complexity of kidney architecture. Recently, three-dimensional cultures of human kidney tubule cells, called tubuloids, and microfluidic platforms that support dynamic flow have provided new opportunities to model kidney function in vitro.

Methods: We developed a human distal tubuloid-on-a-chip model by integrating primary human tubuloid cells with the OrganoPlate® microfluidic culture system. Tubuloids were seeded to form three-dimensional tubular structures adjacent to a collagen type I matrix and exposed to alternating flow. Differentiation into a distal nephron phenotype was evaluated using quantitative polymerase chain reaction, immunohistochemistry, and barrier integrity assays. Functional sodium and water transport were investigated using radiolabeled sodium uptake, transepithelial resistance, dextran diffusion, and dome formation. Pharmacological studies were conducted using trimethoprim, an inhibitor of epithelial sodium channel activity.

Results: Three-dimensional cultures under flow conditions showed increased expression of markers specific to distal nephron segments compared to conventional two-dimensional cultures. Immunostaining confirmed the formation of a highly polarized epithelium with apical and basolateral localization of key transport proteins. The tubules formed a leak-tight barrier, demonstrated by reduced dextran permeability and elevated transepithelial resistance. Radiolabeled assays revealed active sodium transport driven by apical epithelial sodium channels and basolateral sodium-potassium ATPase. Water transport accompanied sodium movement, indicated by dome formation beneath the epithelial layer. Exposure to trimethoprim reduced sodium uptake and dome formation, confirming functional responsiveness of the model.

Conclusions: This distal tubuloid-on-a-chip system reproduces key structural and functional features of the human distal nephron. It enables the study of salt and water transport in primary human tubule cells under controlled microfluidic flow. The model provides a physiologically relevant and scalable platform for investigating kidney physiology, pathology, and pharmacological responses, with potential applications in drug discovery and toxicity testing.

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来源期刊
Kidney360
Kidney360 UROLOGY & NEPHROLOGY-
CiteScore
3.90
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