Culture and analysis of kidney tubuloids and perfused tubuloid cells-on-a-chip
Culture and analysis of kidney tubuloids and perfused tubuloid cells-on-a-chip
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肾小管和灌注小管细胞芯片的培养和分析
DOI：
10.1038/s41596-020-00479-w
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发表时间：
2021-03-05
期刊：
影响因子：
14.8
通讯作者：
Clevers, Hans
中科院分区：
文献类型：
--
作者：
Gijzen, Linda;Yousef Yengej, Fjodor A.;Clevers, Hans
克勒弗斯等人。描述了从组织和尿液中建立肾小管培养物，以及在灌注式微流控多芯片平台中建立肾小管细胞来源的三维小管结构的方法和特征。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。肾小管可由成人肾组织或尿液中的原代细胞形成。小管是一种三维多细胞结构，它概括了小管的功能，已被用于研究感染性、恶性、代谢和遗传性疾病。为了更好地代表活体肾脏，它们可以被集成到一个芯片上的器官系统中，该系统具有更具生理性的管状结构，允许与来自其他器官的血管系统或上皮和间充质细胞流动和相互作用。在这里，我们描述了一种从组织和尿液(1-3周)建立管样培养的详细方案，以及在灌流微流控多芯片平台(7 D)中产生和表征管样细胞来源的三维管状结构的详细方案。这两个系统的结合产生了一个强大的体外工具，可以更好地概括具有供体特有特性的肾小管的复杂性。
Clevers et al. describe the establishment of tubuloid cultures from tissue and urine, as well as the generation and characterization of tubuloid cell-derived 3D tubular structures in a perfused microfluidic multi-chip platform.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.