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Pro�le - Non Viusal Area

Drug Efficacy and Safety Testing

Two-lineSection Head

Applicationsand Related Platforms

Modeling Human Liver DiseaseThe 3D InSight™ HumanLiver Disease platform enables thestudy of NAFLD induction and inhibition of disease progression from steatosis to NASH and fibrosis. The screening- compatible Akura™ 3D microtissue format, provides maximal endpoint compatibility while optimizing efficiency of microtissues handling processes.

Physiologically and mechanistically relevantin vitro system

Powered by AkuraTM Technology

Healthy

in vivo

in vitro

Steatosis NASH Fibrosis

Albumin H&E Oil Red O CD163 Col I

3D InSightTM Liver Disease Platform

Stimulation of disease

Inhibition of diseaseby compounds

Akura™ 96 assay plate1 tissue per well

Assay endpoints

Histology/IHC

Biomarker analysis

High-content analysis

Gene expression

Biochemical assays

● Treatment with free fatty acid (FFA)LPS, and sugars

● Anti-in�ammatory● Anti-�brotic● Metabolic

Delivery Format

Key

Primary Human Hepatocyte (PHH)

Lipid-loadedHepatocyte

Kup�er cell (KC) Hepatic Stellate Cell (HSC)

Liver Endothelial Cell (LEC)

All critical human liver cells and inducers needed to recapitulate the progression of human liver disease, from steatosis to steatohepatitis and fibrosis, are included in the models.

To ensure your success with these complex liver disease models, we provide: ●

expert scientific support ●

detailed assay protocols● the option to bring our models

in-house or outsource assay services to our team of 3D experts

3D InSight™ Human Liver Disease Model

Healthy conditions(control)

Diabetic conditions(lipid loaded)

Cel

lMas

k™ D

eep

Red

Pla

sma

mem

bran

e st

ain

Hoe

chst

333

42 n

ucle

i sta

in |

Nile

Red

lipi

d st

ain

Elafibranor Study with 3D InSight™ NASH Model

3D InSight™Human Liver Disease ModelFor custom applications with non-standard inducers.

3D InSight™Human Liver NASH ModelFor applications that involve the study of the full disease progression from steatosis to NASH and fibrosis. Includes FFA, LPS, and specialized media to mimic diabetic conditions

3D InSight™Human Liver Steatosis ModelIncludes FFA for lipid loading of hepatocytes and specialized media to mimic diabetic conditions.

3D InSight™Human Liver Fibrosis ModelFor applications that require induction of fibrotic scarring in liver tissue. Includes TGF-β and specialized media.

Recapitulating human liver disease with advanced human liver co-culture models

Fibrotic phenotype observed at day 10 of treatment

Elafibranor treatment reduces inflammation

Elafibranor treatment does not affect cell viability of 3D model

Con

trol

FFA

FFA

+ LP

SFF

A+L

PS+

Alk

5i

Col I Col I Col III Col IV α-SMA Collagen Fibrils(count)

(77)

(177)

(699)

(64)

Lipid loading

Pro-fibroticstimuli

Inflammatorystimuli

Fataccumulation

Liverinflammation

Liverscarring

Albumin (PHH) CD31 (LECs)

Con

trol

FFA

+LPS

CD68 (KCs) Vimentin (HSCs)

Model contains relevant cell types to replicate disease states

The model is composed of heathy human primary liver cell types needed for the study of steatosis, NASH, and fibrosis. Treatment with free fatty acids (FFA) and NASH stimuli does not affect cell composition. Immunohistochemistry at day 10 shows expression of characteristic markers for PHHs (Albumin), LECs (CD31), KCs (CD68) and HSCs (Vimentin).

Treatment with FFA induces lipid loading as shown by Nile Red Staining (green). Luminex analysis of secreted cytokines and chemokines after 5 days of treatment with NASH stimuli reflects release of proinflammatory markers

Elafibranor (PPARα/δ agonist) treatment reduces secretion of inflammatory cytokines and chemokines. Luminex analysis of secreted cytokines and chemokines at day 5 of treatment. Bars represent average of 4 microtissues +/- SD. *p ≤ 0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001, comparison between FFA+LPS and FFA+LPS+Elafibranor.

The fibrotic phenotype is observed upon treatment with FFA alone and FFA+LPS. Addition of Alk5 inhibitor blocked blocked development of fibrosis upon treatment with FFA+LPS. Overview (left column) and close-ups of immunohistochemistry staining for Col I, Col III, Col IV, and α-SMA. Right column shows visualization of collagen fibers (arrowheads) by Sirius Red staining and polarized light. Black pixel count (brackets) was determined as the parameter for fibrosis.

0

5

1 0

1 5

Control

TNF-α

TNF-

α (p

g/m

L)

FFA NASH

**

0

5 00

1 00 0

1 50 0

2 00 0

Control FFA NASH

IL-8, MCP-1, MIP-1α IL-8/CXCL8

IL-8

, MC

P-1,

MIP

-1α

(pg/

mL)

MCP-1/CCL2MIP-1α/CCL3

0

2 0

4 0

6 0

8 0

1 00 IL-6

IL-6

(pg/

mL)

Control FFA NASH

***

0

2

4

6

8

1 0

1

Day 2 Day 5 Day 7LHD

rele

ase

(fold

cha

nge

over

veh

icle

con

trol) LDH (day 2, 5, and 7)

V.CtrlElafibranor conc (μM)

1.3 4.0 12.7 40.0 V.CtrlElafibranor conc (μM)

1.3 4.0 12.7 40.0 V.CtrlElafibranor conc (μM)

1.3 4.0 12.7 40.01 1 0 1 00

0

2 0

4 0

6 0

8 0

1 00

IC50 > 40 μmol/L

Cel

l via

bilit

y(%

of c

ontro

l)

ATP (day 7)

Elafibranor conc (μM)

0

2 0

4 0

6 0

8 0

1 00

IL-6

(pg/

mL)

IL- 6

Ctrl FFA+LPS FFA+LPS

Elafibranor conc (μM)1.0 2.0 5.0 10.0 20.0 40.0

IL- 8

MIP

-1α

(pg/

mL)

0

2 00

4 00

6 00

8 00

1 00 0

Ctrl FFA+LPS FFA+LPS

Elafibranor conc (μM)1.0 2.0 5.0 10.0 20.0 40.0

MIP-1α/CCL3

0

5

1 0

1 5

Ctrl FFA+LPS FFA+LPS

Elafibranor conc (μM)1.0 2.0 5.0 10.0 20.0 40.0

TNF-

α (p

g/m

L)

TNF-α

0

5 00

1 00 0

1 50 0

Ctrl FFA+LPS FFA+LPS

Elafibranor conc (μM)1.0 2.0 5.0 10.0 20.0 40.0

MIP

-1α

(pg/

mL)

MCP-1/CCL2

0

5 00

1 00 0

1 50 0

2 00 0

Ctrl FFA+LPS FFA+LPS

Elafibranor conc (μM)1.0 2.0 5.0 10.0 20.0 40.0

IL- 8/CXCL8

IL-8

(pg/

mL)

* **

***

*

*

** **

****

***

*

*

*

**

**

****

*** ****

Certified Applications OptionsDrug efficacy screening ● ● ●

Drug mechanism of action ● ●

Combined drug efficacy and toxicity testing ● ●

Mechanism of disease progression ● ●

Contribution of cell types to disease progression ● ●

Testing potential of drugs to cause disease ● ●

● Model for in-house research ● Assay service ● Technical protocol

Related Models and Platforms

3D InSight™ Diabetes Discovery Platform● 3D InSight™ Diabetes Type II Model● 3D InSight™ Diabetes Type I Model

3D InSight™ Liver Toxicology Platform● 3D InSight™ Human Liver Models● 3D InSight™ Animal Liver Models

Our liver disease platform is built around a precisely engineered co-culture microtissue composed of healthy human hepatocytes, liver endothelial cells, hepatic stellate cells, and Kupffer cells. To enable study of liver disease and drug discovery, we offer different disease models that include our recommended media, inducers, and protocols with instructions on how to induce the desired disease state: steatosis, NASH, and/or Fibrosis. If you are testing alternative inducers or would like to use the healthy liver co-culture model for other applications. Talk to us about your research objectives and we can work together on an appropriate plan and ensure your success with our liver disease models.

Physiologically Relevant Disease Model3D microtissue co-culture of PHHs, KCs, and LECs, and HSCs