liver disease platform poster sm - insphero · 2019-06-20 · study of nafld induction and...
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Pro�le - Non Viusal Area
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