Three-dimensional spheroid culture induces apical-basal polarity and the original characteristics of immortalized human renal proximal tubule epithelial cells
Three-dimensional spheroid culture induces apical-basal polarity and the original characteristics of immortalized human renal proximal tubule epithelial cells
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DOI：
10.1016/j.yexcr.2021.112630
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发表时间：
2021-05-13
影响因子：
3.7
通讯作者：
Asai, Kiyofumi
中科院分区：
文献类型：
--
作者：
Mizuguchi, Ken;Aoki, Hiromasa;Asai, Kiyofumi
近端小管是肾脏的一部分，通过吸收氨基酸、葡萄糖、水和钠、钾和碳酸氢盐等离子来维持血液稳态。近端小管功能障碍与许多肾脏疾病的发病机制有关。肾近端小管上皮细胞（rptec）负责近端小管的主要功能。因此，利用rptec进行体外实验将大大提高我们对肾元生理和病理生物学的认识。最好使用永生化细胞系，如人类肾-2 （HK-2）细胞，因为它们来源于人类并能无限期地保持生长。然而，组织特异性RPTEC表型，包括尖基极化，在传统的二维培养方法中经常丢失，部分原因是微环境缺陷。为了克服这一限制，我们开发了一种使用细胞外基质的HK-2细胞三维（3D）球体培养方法。HK-2球体在三维培养中形成具有细胞极性的小管状结构，并显示出明显恢复的Na转运功能。HK-2细胞的3D培养也增加了肾脏发育相关基因的表达，包括WNT9B。利用HK-2球体建立人肾小管模型将大大提高我们对肾脏生理和病理生物学的认识。
The proximal tubules, which are part of the kidney, maintain blood homeostasis by absorbing amino acids, glucose, water, and ions such as sodium (Na), potassium, and bicarbonate. Proximal tubule dysfunction is associated with the pathogenesis of many kidney diseases. Renal proximal tubular epithelial cells (RPTECs) are responsible for the main functions of the proximal tubules. Therefore, in vitro experiments using RPTECs would greatly enhance our understanding of nephron physiology and pathobiology. It is preferable to use immortalized cell lines, such as human kidney-2 (HK-2) cells, because they are derived from humans and maintain growth indefinitely. However, tissue-specific RPTEC phenotypes, including apical-basal polarization, are frequently lost in conventional two-dimensional culture methods in part due to microenvironmental deficiencies. To overcome this limitation, we developed a three-dimensional (3D) spheroid culture method for HK-2 cells using an extracellular matrix. HK-2 spheroids in 3D culture formed a tubule-like architecture with cellular polarity and showed markedly restored Na transport function. 3D culture of HK-2 cells also increased expression of kidney development-related genes, including WNT9B. Models of human renal tubules using HK-2 spheroids will greatly improve our understanding of the physiology and pathobiology of the kidney.