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Kidney Week

Abstract: TH-OR033

An Immunoprotected Bioreactor for Implanted Renal Cell Therapy

Session Information

  • Bioengineering
    November 07, 2019 | Location: 146 A/B, Walter E. Washington Convention Center
    Abstract Time: 04:54 PM - 05:06 PM

Category: Bioengineering

  • 300 Bioengineering

Authors

  • Gologorsky, Rebecca C., University of California, San Francisco, San Francisco, California, United States
  • Kim, Eun jung, University of California, San Francisco, San Francisco, California, United States
  • Moyer, Jarrett, University of California, San Francisco, San Francisco, California, United States
  • Santandreu, Ana, University of California, San Francisco, San Francisco, California, United States
  • Blaha, Charles, University of California, San Francisco, San Francisco, California, United States
  • Wright, Nathan, University of California, San Francisco, San Francisco, California, United States
  • Chui, Benjamin W., University of California, San Francisco, San Francisco, California, United States
  • Brakeman, Paul R., University of California, San Francisco, San Francisco, California, United States
  • Vartanian, Shant M., University of California, San Francisco, San Francisco, California, United States
  • Humes, H. David, University of Michigan Medical School, Ann Arbor, Michigan, United States
  • Fissell, William Henry, Vanderbilt University, Nashville, Tennessee, United States
  • Roy, Shuvo, University of California, San Francisco, San Francisco, California, United States
Background

An implantable bioartificial kidney will use a bioreactor containing human cells to provide renal cell therapy. The bioreactor must provide an immunoprotected environment for renal tubule cells (RTC) that obviates the need for immunosuppression. Furthermore, RTC differentiation and functionality must be maintained in vivo.

Methods

We developed an immunoprotection chamber using silicon nanopore membranes (SNM) with sub-10nm wide slit pores and evaluated primary human renal epithelial cell viability and functionality in a benchtop model and in vivo. For the benchtop, we created a stacked dual-chamber vessel, with human RTC in the inferior chamber isolated by SNM. After exposure to 500ng/mL TNF-α, RTC were evaluated for viability and monolayer integrity using cell viability assay, transepithelial electrical resistance, and immunohistochemistry. Thereafter, we implanted an analogous bioreactor in a pig without systemic anticoagulation, with the device perifused by connections to the carotid artery and jugular vein. A static in vitro control was used for comparison. The device was explanted after three days for RTC evaluation. ELISA and quantitative PCR (qPCR) were used to examine RTC-specific markers that are surrogates for RTC functionality.

Results

Benchtop testing showed that isolated RTC were confluent and >90% viable, whereas cells directly exposed to TNF-α were nonviable. In vivo, there were no thrombi in the device. Cells were confluent with >90% viability and maintained intercellular interaction/signal transduction via tight junctions/diffuse expression of Zona Occludens-1 protein similar to static control. Implanted RTC had low NAGexpression at <10% the rate of positive control indicating minimal damage to RTC while implanted. Expression of AQP1, 1a Hydroxylase, and NHE3 was up-regulated in implanted cells and twofold higher than static controls, suggesting greater functionality of implanted cells.

Conclusion

We present a kidney bioreactor tested in vivo that sustains implanted human renal cells in a xenogeneic environment without systemic anticoagulation. These data demonstrate the promise of a SNM-based bioreactor for use in implantable bioartificial kidney without immunosuppression.

Funding

  • Other NIH Support