Abstract: TH-PO321

Synthesized Basement Membrane Substratum Provided Cultured Renal Tubular Cells with Scaffold upon Which They Aggressively Developed Filolpodia/Lamellipodia Needed for Cell Motility

Session Information

Category: Acute Kidney Injury

  • 002 AKI: Repair and Regeneration

Authors

  • Inui, Kiyoko, Division of Nephrology, Department of Medicine, Showa University Fujigaoka Hospital, Yokohama, Japan
  • Morita, Hiroyuki, Aichi Medical University School of Medicine, Nagakute, Japan
  • Inoue, Yoshihiko, Division of Nephrology, Department of Medicine, Showa University Fujigaoka Hospital, Yokohama, Japan
  • Kawashima, Eri, Division of Nephrology, Department of Medicine, Showa University Fujigaoka Hospital, Yokohama, Japan
  • Koiwa, Fumihiko, Division of Nephrology, Department of Medicine, Showa University Fujigaoka Hospital, Yokohama, Japan
  • Mochitate, Katsumi, National Institute for Environmental Studies, Tsukuba, Japan
  • Yoshimura, Ashio, Division of Nephrology, Department of Medicine, Showa University Fujigaoka Hospital, Yokohama, Japan
Background

Little is known about the ultrastructure of renal tubular epithelium in culture. We previously reported imaging obtained by the use of scanning electron microscopy (SEM) for cultured cell allowed curious aspects of cellular morphology for discussion. In this study, we aimed to capture SEM images of synthesized basement membrane (sBM) we developed and see how it created conditions favorable for cellular proliferation.

Methods

The collagenous substrata, “fib”, were prepared on 2 chamber culture dishes interconnected with pores. Type I collagen solution was cast on the second chamber, polymerized, air-dried and used as fib. SV40-T2 cells were seeded on fib, and cocultured with EHS tumor matrigel in the first chamber. During a week of culture, a lamina densa structure formed beneath the cells. The cells were removed and used as sBM substrata.

Results

In subculturing the cells, rattus norvegicus kidney tubular epithelialium, NRK-52E, attached fib substrata (Figure), formed lamellipodia, assembled, and proliferated until they became confluent. Application of sBM substrata increased the growth rate of NRK-52 3-fold. SEM images disclosed formation of filopodia / lamellipodia and cell flattening were much more aggresive with sBM substrata.

Conclusion

In vitro, sBM substratum provided NRK-52E cell with scaffold upon which it easily developed cellular structures needed for cell motility and proliferation.