Fig. 1
From: Proximal tubule-on-chip as a model for predicting cation transport and drug transporter dynamics
Flow shear stress influences drug transporter transcriptomic expression and endogenous metabolite production. (A) Protocol used for RPTEC/TERT1 cell culture in proximal tubule-on-chip in the present study. (B-C) Fold changes regarding the mRNA expressions of proximal tubule ABC and SLC transporters of pharmacological relevance. Bar plots show relative expression levels of RPTEC/TERT1 mRNA transporter expressions using mono-channel device where cells are exposed to different flow rates, 5 µL/min (0,0045 dyn/cm2, N = 6), 10 µL/min (0,009 dyn/cm2, N = 11) and 20 µL/min (0,02 dyn/cm2, N = 4). It is worth mentioning that for SLC22A4, SLC22A5 and SLC47A2, only three independent replicas (N = 3) were performed. Expression levels were normalized to the internal control GAPDH gene and are presented as fold changes relative to rocker-based perfusion conditions (N = 5, dotted line). Error bars represent standard deviations. Statistical significances were conducted using one-way ANOVA analysis, asterisks (*) indicate statistically significant differences compared to rocker-based perfusion condition (*p-value < 0.05, **p-value < 0.01, ***p-value < 0.001). (D) Targeted search for 33 endogenous metabolites out of 103 initially considered (see main text) by LC–MS/MS on extracellular supernatant of RPTEC/TERT1 cells culture in rocker-based perfusion (Rocker) condition or exposed to different flow rates (5, 10 and 20 μL/min). Data were standardized by auto scaling and hierarchically clustered with Ward method and represented by mean of 3 independent replicates. Metabolomic clustering was achieved using the MetaboAnalyst 6.0 Statistical Analysis online tool.
