Supplementary Information

Liver Injury-on-a-Chip: Microfluidic Co-Cultures with Integrated Biosensors for

Monitoring Liver Cell Signaling During Injury

Qing Zhou#1, Dipali Patel#1, Timothy Kwa1, Amranul Haque1, Zimple Matharu1, Gulnaz

Stybayeva1, Yandong Gao1, Anna Mae Diehl2, Alexander Revzin1

Affiliation:

1 Department of Biomedical Engineering, University of California, Davis, 451 Health Sciences,

Davis, CA, USA.

2 Division of Gastroenterology, Department of Medicine, Duke University, 595 LaSalle Street,

Snyderman Building, Suite 1073, Durham, NC

* Correspondence and requests for materials should be addressed to:

Alexander Revzin (arevzin@ucdavis.edu)

Electronic Supplementary Material (ESI) for Lab on a Chip.This journal is © The Royal Society of Chemistry 2015

Supplementary Figure Legends

Supplementary Figure 1. Maintenance of epithelial phenotype of hepatocyte and quiescent

phenotype of stellate cells inside microchambers. Immunostaining images of epithelia marker

(Albumin, green) and quiescent stellate cell marker (GFAP, red) in hepatocytes and stellate cells,

respectively. Cells were cultured for 7 days inside microchambers and standard tissue culture

plates pre-coated with collagen type I. DAPI was used to stain nucleus. Scale bar: 50 µm. GFAP:

Glial Fibrillary Acidic Protein.

Supplementary Figure 2. Alcohol injured hepatocytes activates neighboring stellate cells.

Expression of α-smooth muscle actin (α-SMA, green) in stellate cells cultured for 48 h inside

microchambers next to healthy hepatocytes [EtOH (-)] and hepatocytes injured for 48 h with

alcohol [EtOH (+)]. DAPI (blue) was used to stain nucleus.

Supplementary Figure 3. Layout of the three-layer sensing device containing micropatterned

electrodes on the substrate and the reconfigurable PDMS top layers.

Supplementary Figure 4. Proof -of-concept experiments to demonstrate the independent

operation of electrode columns in sensing channels. (A) Alexa 546 solution was infused in all

three sensing channels (a,b,c). (B-D) Sensing channels were selectively actuated via vacuum

control. Corresponding graphs represent fluorescence intensity of acquired signals.

Supplementary Figure 5. Selective stimulation of sensing chambers with recombinant TGF-.

(a-c) 100ng/ml recombinant TGF-β was infused into two cell culture chambers, followed by

sequential exposure of electrode column a, column b and column c, with one after another. The

activation of designated electrode columns was reflected by the suppression of SWV curves in

panel a, b and c. The black arrows indicate the appliance of negative pressure in correspondent

control chamber.

Supplementary Figure 6. Numerical simulation of the cytokine diffusion and surface binding

process. (a-b) Simulated signal (Blue) under assumed secretion rate (σH= 0.00576pg/h; σs=

0.00159pg/h) and real experimental data (Red) plotted over time for comparison purpose. (c)

Concentration distribution of cytokine (TGF-β1) in the microfluidic chamber predicted by the

computational model.

Supplementary Figure 1

Supplementary Figure 2

Supplementary Figure 3

Supplementary Figure 4

Supplementary Figure 5

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Supplementary Figure 6

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(a) (b)Hepatocytes: σH=0.00576pg/h

Stellate Cells: σs=0.00159pg/h

(c)