Abstract: FR-PO992

3-D Kidney-on-Chip Platform for Quantitative Screening of Podocyte Structural Integrity

Session Information

Category: Bioengineering and Informatics

  • 101 Bioengineering and Informatics

Authors

  • Bhattacharya, Smiti, Columbia University, New York, New York, United States
  • Ron, Amit, Columbia University , New York, Nevada, United States
  • Calizo, Rhodora C., Icahn School of Medicine at Mount Sinai, New York, New York, United States
  • Wiener, Robert, Icahn School of Medicine at Mount Sinai, New York, New York, United States
  • He, John C., Mount Sinai School of Medicine, New York, New York, United States
  • Iyengar, Ravi, Icahn School of Medicine at Mount Sinai, New York, New York, United States
  • Hone, James C, Columbia University, New York, New York, United States
  • Azeloglu, Evren U., Icahn School of Medicine at Mount Sinai, New York, New York, United States
Background

Podocytes have an intricate in vivo morphology that is critical for their physiological function. Under disease conditions or upon isolation and culture, podocytes dedifferentiate and lose their specialized morphology, which prevents the use of primary or immortalized podocyte lines as translatable in vitro models of chronic kidney disease. In order to systematically study podocyte mechanobiology and drug response, we developed an in vitro culture system that utilizes microfabricated 3-D biochips to mechanically induce formation of fine peripheral processes in podocytes.

Methods

We microfabricated 3-D biochips using photolithography. We developed biochips of varying size formats from standard 96-well plates to 25-mm coverslips. Immortalized human podocytes were plated on 3-D biochips and differentiated for five days at 37 degrees Centigrade. Transcriptomic expression of differentiation markers was quantified using RT-PCR. Spatial localization and protein expression were quantified using immunofluorescence. Spatial biomechanics were quantified using atomic force microscope elastography.

Results

Podocytes in 3-D biochips displayed significant upregulation of a wide range of genes associated with the differentiated phenotype. The peripheral processes were selectively enriched for slit diaphragm components nephrin, podocin and neph1 as well as crosslinked actin bundles. Micropatterned podocytes exhibited heterogeneous biomechanical properties with significantly increased elastic modulus in peripheral processes. This spatial phenotype was lost when cells were treated with known nephrotoxic drugs or inhibitors of cytoskeletal integrity. When we looked at phenotypic signatures of protein localization, focal adhesion maturation and cytoskeletal integrity, podocytes in biochips showed reduced cell-to-cell variability and high reproducibility compared to those on unpatterned glass surfaces.

Conclusion

We developed a 3-D kidney-on-chip system that provides a quantitative, high-throughput in vitro platform for studying podocyte morphology, biomechanics and drug response with high reproducibility.

Funding

  • NIDDK Support