Abstract: TH-OR139
A High-Throughput Microfluidic Model of Proximal Tubule with Active Transport Function
Session Information
- Translational Research Innovations in Kidney Organoid and Bioengineered Model Systems
October 25, 2018 | Location: 4, San Diego Convention Center
Abstract Time: 06:06 PM - 06:18 PM
Category: Bioengineering
- 300 Bioengineering
Authors
- Vedula, Else M., Draper, Cambridge, Massachusetts, United States
- Haggerty, Timothy J., Draper, Cambridge, Massachusetts, United States
- Charest, Joseph L., Draper, Cambridge, Massachusetts, United States
Background
Models with kidney-specific function in vitro and high-throughput capability will speed kidney disease drug development. Microfluidic models control flow to cultured cells to generate tissue with kidney-specific function. However, high throughput is required for experiments accommodating many drug doses, assays, and biological variables. Microfluidic models have yet to achieve high throughput.
Methods
We developed an in vitro kidney model in a high-throughput microfluidic platform, PREDICT-96. PREDICT-96 co-cultured human renal proximal tubule epithelial cells (hRPTEC) and human microvascular endothelial cells (hMVEC) and characterized them via real-time trans-epithelial electrical resistance (TEER) and a high-content screening (HCS) approach quantifying ZO-1 expression, localization. Primary cilia, polycystin-1 (PC1), and transporter expression were characterized via confocal microscopy. Glucose reabsorption via SGLT2 and organic anion transport via OAT-1 were characterized using real-time imaging and spectrophotometric techniques.
Results
hRPTEC and hMVEC formed polarized tissue with barrier integrity and active, kidney-specific transport function. Flow increased TEER, hRPTEC thickness, primary cilia expression with co-localized PC1, and ZO-1 localization. Drug dosing modulated glucose reabsorption from the filtrate to vascular channel, indicating active kidney-specific function. OAT-1 expression and function were characterized to quantify uptake and excretion functions.
Conclusion
Microfluidic flow enhanced tissue structure/morphology, polarization, and function, resulting in in vitro tissue with kidney-specific function. Integrated TEER sensing and HCS-capable platform architecture supported real-time data collection. Microfluidic flow-conditioned tissue actively responded to drug dosing, indicating the ability of the model to quantify drug effects. The PREDICT-96 kidney model can generate kidney tissue, study renal toxicity, and evaluate disease progression in vitro to speed development of kidney therapeutics.