A novel method for successful induction of interdigitating process formation in conditionally immortalized podocytes from mice, rats, and humans.
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.
['Doi|Kotaro|K|', 'Kimura|Hiroshi|H|', 'Wada|Takehiko|T|', 'Tanaka|Tetsuhiro|T|', 'Hiromura|Keiju|K|', 'Saleem|Moin A|MA|', 'Inagi|Reiko|R|', 'Nangaku|Masaomi|M|', 'Fujii|Teruo|T|']
D000199:Actins D000818:Animals D018929:Cell Culture Techniques D002461:Cell Line, Transformed D022081:Cell Surface Extensions D003599:Cytoskeleton D005786:Gene Expression Regulation D006358:Hot Temperature D006801:Humans D051379:Mice D050199:Podocytes D012333:RNA, Messenger D051381:Rats D013045:Species Specificity D014746:Vimentin
Process formation Immortalized podocytes Confluence Cytoskeleton