IRCCS - Istituto di Ricerche Farmacologiche 'Mario Negri', Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy.
Nanomed Laboratories, Dipartimento di Fisica, Università di Genova, 16146 Genova, Italy.
EBioMedicine. 2018 Jul;33:253-268. doi: 10.1016/j.ebiom.2018.06.005. Epub 2018 Jul 3.
The lack of engineering systems able to faithfully reproduce complex kidney structures in vitro has made it difficult to efficiently model kidney diseases and development. Using polydimethylsiloxane (PDMS) scaffolds and a kidney-derived cell line we developed a system to rapidly engineer custom-made 3D tubules with typical renal epithelial properties. This system was successfully employed to engineer patient-specific tubules, to model polycystic kidney disease (PKD) and test drug efficacy, and to identify a potential new pharmacological treatment. By optimizing our system we constructed functional ureteric bud (UB)-like tubules from human induced pluripotent stem cells (iPSCs), and identified a combination of growth factors that induces budding morphogenesis like embryonic kidneys do. Finally, we applied this assay to investigate budding defects in UB-like tubules derived from a patient with a PAX2 mutation. Our system enables the modeling of human kidney disease and development, drug testing and discovery, and lays the groundwork for engineering anatomically correct kidney tissues in vitro and developing personalized medicine applications.
缺乏能够真实再现体外复杂肾脏结构的工程系统,使得高效地对肾脏疾病和发育进行建模变得困难。我们使用聚二甲基硅氧烷(PDMS)支架和一种肾脏来源的细胞系,开发了一种快速工程定制 3D 管状结构的系统,这些管状结构具有典型的肾上皮特性。该系统成功地用于工程化患者特异性管状结构,模拟多囊肾病(PKD)并测试药物疗效,并确定了一种潜在的新的药理学治疗方法。通过优化我们的系统,我们从人诱导多能干细胞(iPSC)构建了功能性输尿管芽(UB)样管状结构,并确定了一种组合的生长因子,该因子诱导类似于胚胎肾脏的萌芽形态发生。最后,我们应用该检测方法来研究来源于 PAX2 突变患者的 UB 样管状结构中的萌芽缺陷。我们的系统使人类肾脏疾病和发育的建模、药物测试和发现成为可能,并为在体外构建解剖学上正确的肾脏组织和开发个性化医疗应用奠定了基础。