Abstract: PO0495
Modeling Ischemia-Reperfusion in a High-Throughput Tubular/Microvascular Co-Culture-on-a-Chip
Session Information
- Bioengineering: Organoids and Organs-on-a-Chip
November 04, 2021 | Location: On-Demand, Virtual Only
Abstract Time: 10:00 AM - 12:00 PM
Category: Bioengineering
- 300 Bioengineering
Authors
- Shaughnessey, Erin M., Tufts University, Medford, Massachusetts, United States
- Kann, Samuel H., Boston University, Boston, Massachusetts, United States
- Black, Lauren D., Tufts University, Medford, Massachusetts, United States
- Charest, Joseph L., Draper, Cambridge, Massachusetts, United States
- Vedula, Else M., Draper, Cambridge, Massachusetts, United States
Background
Ischemia-reperfusion (IR) is a major cause of acute kidney injury (AKI). IR involves a period of inadequate perfusion that deprives highly metabolic kidney epithelia of nutrient and waste exchange impacting tissue structural integrity.
Methods
We demonstrate the potential of the PREDICT96 (P96) platform as a tool for examining tubule cell responses to IR. The oxygen-impermeable construction and robust pumping capabilities of P96 enable comparison of simulated IR and control conditions on a single culture plate. Here, primary human renal proximal tubule epithelial cells and human microvascular endothelial cells were cultured in adjacent microfluidic channels for 5 days under physiological fluid shear (0.07 Pa) to establish confluent layers. Subsequently, a portion of the tissue replicates underwent simulated IR consisting of 3 days of static conditions followed by 2 days of physiological flow. Transepithelial electrical resistance (TEER) was measured daily, and all tissues were fixed after 10 days in culture for structural characterization via immunofluorescence confocal microscopy.
Results
TEER measurements highlighted a transient increase in barrier function (relative to control tissues) in response to the onset of ischemia, followed by a reduction in integrity over subsequent days that persisted through reperfusion. IR tissues exhibited primary cilia that were on average less abundant but roughly double the length of those exhibited by control tissues. This is consistent with previous observations of cilia lengthening among IR-injured kidney epithelia in vivo. IR tissues also displayed nuclear staining for β-catenin suggestive of a proliferative response to injury.
Conclusion
The described model has significant potential to clarify mechanisms of IR injury in the kidney.
(A) Experimental scheme for simulated IR in P96. (B) Average change in TEER by group relative to baseline readings on day 5 at start of IR treatment. N=12 devices per group. *p<0.05 t-test. (C) Change in tissue morphology following IR treatment.
Funding
- Other U.S. Government Support