A novel method for successful induction of interdigitating process formation in conditionally immortalized podocytes from mice, rats, and humans

Kotaro Doi, Hiroshi Kimura, Takehiko Wada, Tetsuhiro Tanaka, Keiju Hiromura, Moin A Saleem, Reiko Inagi, Masaomi Nangaku, Teruo Fujii

Research output: Contribution to journalArticle (Academic Journal)peer-review

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Abstract

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.

Original languageEnglish
Pages (from-to)47-52
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume570
Early online date14 Jul 2021
DOIs
Publication statusPublished - 17 Sep 2021

Bibliographical note

Funding Information:
We thank Dr. Hidetake Kurihara for providing the rat podocyte cell line 2DNA1D7. This study was partially supported by the Japanese Agency for Medical Research and Development, Tokyo, Japan (Grant No. 18be0304204h0002 ) (to D.K., H.K., T.T., R.I., M.N. and T.F.). The authors declare no conflicts of interest.

Funding Information:
The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Teruo Fujii reports financial support was provided by the Japanese Agency for Medical Research and Development, Tokyo, Japan . Kotaro Doi, Hiroshi Kimura, Tetsuhiro Tanaka, Reiko Inagi, Masaomi Nangaku reports financial support was provided by the Japanese Agency for Medical Research and Development, Tokyo, Japan .

Publisher Copyright:
© 2021 The Authors

Keywords

  • Actins/metabolism
  • Animals
  • Cell Culture Techniques/methods
  • Cell Line, Transformed
  • Cell Surface Extensions/metabolism
  • Cytoskeleton/metabolism
  • Gene Expression Regulation
  • Hot Temperature
  • Humans
  • Mice
  • Podocytes/metabolism
  • RNA, Messenger/genetics
  • Rats
  • Species Specificity
  • Vimentin/metabolism

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