A novel method for successful induction of interdigitating process formation in conditionally immortalized podocytes from mice, rats, and humans
A novel method for successful induction of interdigitating process formation in conditionally immortalized podocytes from mice, rats, and humans
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DOI：
10.1016/j.bbrc.2021.07.029
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发表时间：
2021-07-14
影响因子：
3.1
通讯作者：
Fujii, Teruo
中科院分区：
文献类型：
--
作者：
Doi, Kotaro;Kimura, Hiroshi;Fujii, Teruo
足细胞中突的形成被认为是细胞成熟和正常生理状态的标志。关于足细胞突回缩的分子机制有许多积累的发现；然而，对于促进体外过程形成的积极机制知之甚少，并且大多数关于该主题的先前报告仅限于低密度培养。在这里，我们发现通过在laminin-521（L521）支架上结合血清消耗和Y-27632 ROCK抑制剂，可以在小鼠、大鼠和人类的条件永生化足细胞的100%融合培养中诱导过程形成。我们注意到在这种情况下，波形蛋白丝径向延伸模式的细胞骨架重组和肌动蛋白应激纤维形成的下调。我们还发现，额外标准量的血清、ROCK抑制剂的缺失或支架与层粘连蛋白-511（L511）的轻微不匹配都会阻碍过程的形成。这些发现表明，减少血清、足细胞特异性支架和细胞内信号的结合，以减少ROCK的过表达，是过程形成的必要因素。(c) 2021作者。Elsevier Inc.出版。这是一篇基于CC BY许可（http://creativecommons.org/licenses/by/4.0/）的开放获取文章。
Formation of processes in podocytes is regarded as the hallmark of maturity and normal physical condition for the cell. There are many accumulated findings about molecular mechanisms that cause retraction of podocyte processes; however, there is little knowledge of the positive mechanisms that promote process formation in vitro, and most previous reports about this topic have been limited to low density cultures. Here, we found that process formation can be induced in 100% confluent cultures of conditionally immortalized podocytes in mouse, rat, and human species by combining serum depletion and Y-27632 ROCK inhibitor supplementation on the scaffold of laminin-521(L521). We noted the cytoskeletal reorganization of the radial extension pattern of vimentin filaments and downregulation of actin stress fiber formation under that condition. We also found that additional standard amount of serum, depletion of ROCK inhibitor, or slight mismatch of the scaffold as laminin-511(L511) hinder process formation. These findings suggest that the combination of reduced serum, podocyte-specific scaffold, and intracellular signaling to reduce the overexpression of ROCK are required factors for process formation. (c) 2021 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).