Department of Urology, Urological Science Institute, Yonsei University College of Medicine, Seoul, Republic of Korea.
Department of Urology, National Health Insurance Service Ilsan Hospital, Gyeonggi-do, Republic of Korea.
PLoS One. 2018 Oct 31;13(10):e0206447. doi: 10.1371/journal.pone.0206447. eCollection 2018.
Cell-culture methods that simplify the inherent complexities of the kidney have not sufficiently reproduced its true characteristics. Although reports indicate that organoid methodology surpasses traditional cell culture in terms of reproducing the nature of organs, the study of human kidney organoids have been confined to pluripotent stem cells. Furthermore, it has not yet progressed beyond the developmental state of embryonic kidney even after complicate additional differentiation processes. We here describe the kidney organotypic culture method that uses adult whole kidney tissues but mainly differentiates into tubular cells. This model was validated based on the retention of key kidney organotypic-specific features: 1) expression of Tamm-Horsfall protein; 2) dome-like organoid configurations, implying directed transport of solutes and water influx; and 3) organoid expression of neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1) in response to nephrotoxic injury (i.e., gentamicin and cisplatin exposure). This 3D-structured organoid prototype of the human renal tubule may have applications in developing patient-specific treatments for kidney diseases.
细胞培养方法虽然简化了肾脏固有的复杂性,但尚未充分再现其真正的特征。虽然有报道称类器官方法在再现器官本质方面优于传统细胞培养,但对人类肾脏类器官的研究仅限于多能干细胞。此外,即使经过复杂的额外分化过程,其也尚未超越胚胎肾脏的发育状态。我们在这里描述了一种使用成人整个肾脏组织但主要分化为肾小管细胞的肾脏器官培养方法。该模型基于保留关键的肾脏器官特异性特征进行了验证:1)Tamm-Horsfall 蛋白的表达;2)穹顶样类器官结构,暗示溶质的定向转运和水流入;3)类器官表达中性粒细胞明胶酶相关脂质运载蛋白 (NGAL) 和肾损伤分子-1 (KIM-1) 以响应肾毒性损伤(即庆大霉素和顺铂暴露)。这种人类肾小管的 3D 结构原型类器官可能在开发针对肾脏疾病的患者特异性治疗方法方面具有应用价值。