Roye Yasmin, Musah Samira
Department of Biomedical Engineering, Pratt School of Engineering, Duke University.
Department of Biomedical Engineering, Pratt School of Engineering, Duke University; Department of Medicine, Division of Nephrology, Duke University School of Medicine; Department of Cell Biology, Duke University; Center for Biomolecular and Tissue Engineering, Duke University;
J Vis Exp. 2022 Nov 4(189). doi: 10.3791/63821.
Chronic kidney disease (CKD) affects 15% of the U.S. adult population, but the establishment of targeted therapies has been limited by the lack of functional models that can accurately predict human biological responses and nephrotoxicity. Advancements in kidney precision medicine could help overcome these limitations. However, previously established in vitro models of the human kidney glomerulus-the primary site for blood filtration and a key target of many diseases and drug toxicities-typically employ heterogeneous cell populations with limited functional characteristics and unmatched genetic backgrounds. These characteristics significantly limit their application for patient-specific disease modeling and therapeutic discovery. This paper presents a protocol that integrates human induced pluripotent stem (iPS) cell-derived glomerular epithelium (podocytes) and vascular endothelium from a single patient to engineer an isogenic and vascularized microfluidic kidney glomerulus chip. The resulting glomerulus chip is comprised of stem cell-derived endothelial and epithelial cell layers that express lineage-specific markers, produce basement membrane proteins, and form a tissue-tissue interface resembling the kidney's glomerular filtration barrier. The engineered glomerulus chip selectively filters molecules and recapitulates drug-induced kidney injury. The ability to reconstitute the structure and function of the kidney glomerulus using isogenic cell types creates the opportunity to model kidney disease with patient specificity and advance the utility of organs-on-chips for kidney precision medicine and related applications.
慢性肾脏病（CKD）影响着15%的美国成年人口，但由于缺乏能够准确预测人类生物学反应和肾毒性的功能模型，靶向治疗的建立受到了限制。肾脏精准医学的进展有助于克服这些限制。然而，此前建立的人类肾小球体外模型——血液过滤的主要部位以及许多疾病和药物毒性的关键靶点——通常采用功能特性有限且基因背景不匹配的异质细胞群体。这些特性显著限制了它们在患者特异性疾病建模和治疗发现中的应用。本文介绍了一种方案，该方案整合了来自单一患者的人类诱导多能干细胞（iPS）衍生的肾小球上皮细胞（足细胞）和血管内皮细胞，以构建一个同基因且血管化的微流控肾小球芯片。由此产生的肾小球芯片由干细胞衍生的内皮细胞层和上皮细胞层组成，这些细胞层表达谱系特异性标志物，产生基底膜蛋白，并形成类似于肾脏肾小球滤过屏障的组织-组织界面。该工程化的肾小球芯片能够选择性地过滤分子，并重现药物诱导的肾损伤。使用同基因细胞类型重建肾小球结构和功能的能力为以患者特异性方式模拟肾脏疾病创造了机会，并提高了芯片器官在肾脏精准医学及相关应用中的效用。
