Abstract: FR-PO993

Biomimetic Microenvironment Promotes In Vivo-Like Phenotype of Conditionally Immortalized Podocytes

Session Information

Category: Bioengineering and Informatics

  • 101 Bioengineering and Informatics

Authors

  • Ishahak, Matthew, University of Miami, Miami, Florida, United States
  • Jones, Ellery, University of Miami, Miami, Florida, United States
  • Fornoni, Alessia, University of Miami, Miami, Florida, United States
  • Agarwal, Ashutosh, University of Miami, Miami, Florida, United States
Background

Microenvironmental cues are integral in providing signals that modulate podocyte development and function. However, standard culture conditions fail to recapitulate the chemical, physical, and architectural cues presented by the glomerular basement membrane (GBM). We report the effect of an engineered GBM-mimic hydrogel on conditionally immortalized human podocyte cultures.

Methods

A hydrogel was developed through the enzymatic crosslinking of denatured collagen to serve as a GBM-mimic. 3D grooves were micromolded into the surface of the biomimetic hydrogel. Mechanical properties of the GBM-mimic hydrogel were characterized using a rheometer fitted with a metal parallel plate in oscillation mode. Conditionally immortalized human podocytes were cultured on various substrates in permissive conditions. Once cells reached ~90% confluence, they were moved to non-permissive conditions and cultured for 14 days to allow for differentiation. Phenotypic features were assessed by fluorescent imaging and scanning electron microscopy (SEM).

Results

A GBM-mimic hydrogel that recapitulates the protein composition, mechanical properties, and 3D surface topography of the GBM was successfully fabricated. The stiffness of the hydrogel (E=5.43±0.69 kPa) was much closer to the physiological stiffness of the glomerulus than that of other cell culture substrates (Fig 1A). SEM revealed the formation of processes branching from the main cell body of podocytes grown on the GBM-mimic hydrogels (Fig 1B).

Conclusion

These results demonstrate that microenvironmental cues induce in-vivo like podocyte morphology. Current studies are focused on incorporating additional GBM components (e.g. laminin) and dynamic physiological phenomenon (e.g. fluid flow, pressure differential, and protein gradient). Ultimately, an integrated platform that maintains and measures long term glomerular filtration will be developed to serve as a powerful addition to the toolkit for studying kidney disease and developing therapeutics.