BIOMIMESYS® range are hyaluronan based hydroscaffold developed to overcome the 2D flat culture limitations by recreating an in vivo-like physiology within the in vitro environment.
BIOMIMESYS®Liver scaffold is made of RGDS and galactosamine-grafted Hyaluronic acid, Adipic acid dihydrazide crosslinker and extracellular matrix (ECM) proteins (collagen type I and collagen type IV) to mimic liver-ECM composition.
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BIOMIMESYS® Liver, a 3D cell culture model for maintaining and promoting hepatocytes functions for metabolism and toxicity studies
1. Human HepatocytesHepG2
BIOMIMESYS® Liver has been tested on HepG2 and on cryopreserved Human Hepatocytes. It makes 3D cell culture easy and provides a robust in
vitro and reliable model for metabolism & toxicological studies.
To know more about BIOMIMESYS®Liver visit our website: www.biomimesys.com
+33(0) 769 999 137; hello@biomimesys.com
BIOMIMESYS®Liver is ready to use and compatible with all analytical technologies
• Available in a ready-to-use format (96 well plates) it enables the culture of hepatocytes under physiological conditions
that are representative of the microenvironment found in liver tissue(2).
• Hepatocytes are simply seeded on top of the hydroscaffold and placed in the incubator.
• The media can be refreshed easily by pipetting. Being transparent makes it suitable for microscopy
(immunofluorescence, bright field) and use in plate readers (OD, fluorescence & luminescence).
• Thanks to its mechanical properties, the hydroscaffold can be easily handled with fine forceps and proteins, nucleic
acid can be extracted by directly adding the lysis buffer to the hydrogel, due to its high porosity.
BIOMIMESYS®Liver is physiological
BIOMIMESYS® range are hyaluronan based hydroscaffold developed to overcome the 2D flat
culture limitations by recreating an in vivo-like physiology within the in vitro environment.
In comparison to in vivo
decellularized liver tissue
SEM observation of a human decellularized liver
tissue (from Mazza et al. 2015(1) )
SEM observation of a BIOMIMESYS®Liver section, highlighting
the collagen chain, (artificially coloured)
Biomimetic structure
References:
(1) Decellularized human liver as a natural 3D-scaffold for liver bioengineering and transplantation. Mazza G. et al. Sci Rep.7; 5:13079, 2015.
(2) Decellularization and cell seeding of whole liver biologic scaffolds composed of extracellular matrix. Faulk D. et al. J Clin Exp Hepatol. 5; 1:69-80, 2015.
BIOMIMESYS®Liver hydroscaffold is made of
RGDS and galactosamine-grafted Hyaluronic
acid, Adipic acid dihydrazide crosslinker and
extracellular matrix (ECM) proteins (collagen
type I and collagen type IV) to mimic liver-
ECM composition.
0
50
100
150
Day 7 Day 14 Day 21 Day 28
Diameter(µm)
HepG2 spheroids sizes
100µm400µm
100µm
Alive
Dead
Day 28
Viability and Growth
Albumin Secretion
• HepG2 form spheroids with an average diameter of 80 to 100 µm after
one month of cultivation.
• The amount of albumin secreted by
HepG2 is 20 to 30 times higher in
BIOMIMESYS®Liver compared to 2D.
• Cryopreserved human hepatocytes form small
aggregates, 40 to 80 µm, which have a very
good viability until day14.
Key genes expression levels
Toxicity study
• Cryopreserved human
hepatocytes grown in
BIOMIMESYS®Liver represent
model for routine drug testing in
96-well format.
n=3
0
20
40
60
80
100
120
Day 7 Day 14
Diameter(µm)
Human Hepatocytes spheroids sizes
donor 1 donor 2
3D
Day 7
2D
100µm
25µm 25µm
Day 21
100µm MRP-2
Biliary Canaliculi Formation
• MRP2 expression, its colocalization
with actin and CFDA fluorescence
confirms the presence of active biliary
canaliculi in HepG2 during 3 weeks.
CYP activities
• Basal and induced CYP1A1/A2, CYP2B6 and CYP3A4 activities are higher in
BIOMIMESYS®Liver compared to 2D collagen sandwich.
Basal activity 2D vs 3D Basal and induced activities in 3D
0
50
100
150
200
CYP1A1/A2
2D 3D
Acetaminophen/100ngof
DNA
0
500
1000
1500
2000
CYP3A4
6β-OH-Testosterone
(nM)/100ngofDNA
0
20
40
60
80
CYP2B6
OH-Bupropion
(nM)/100ngofDNA
0
1000
2000
3000
CYP1A1/A2
Basal Induced
***
Acetaminophen/100ngof
DNA
0
1000
2000
3000
4000
5000
CYP3A4
***
6β-OH-Testosterone
(nM)/100ngofDNA
0
100
200
300
400
500
600
CYP2B6
***
OH-Bupropion
(nM)/100ngofDNA
Day 7
Viability and Growth
100µm
Day 7
100µm
Day 14
400µm
100µm 100µm
• HepG2 grown in BIOMIMESYS®Liver
allow acute and chronic dose (3 doses)
drug exposure. -10%
10%
30%
50%
70%
90%
110%
130%
0,01 0,2 4 80
Viability(ATPcontent)
Chlorpromazine (µM)
BIOMIMESYS®Hepatocyte
-10%
10%
30%
50%
70%
90%
110%
130%
0,1 2 40 800
Amiodarone (µM)
BIOMIMESYS®Liver, a 3D cell culture model for maintaining and
promoting hepatocytes functions for metabolism and toxicity studies
Day 14
400µm
25µm
Day 21
Actin
MRP2
Day 14
CFDA
100µm
Toxicity study
Day 7 Day 8 Day 9 Day 10
1 dose
1st dose 2nd dose 3rd dose
Viability
Viability
Treatment 1 dose : Day 7 - Viability analysis : Day 8
• Better expression of phase I & II enzymes and transporters genes in 3D
Day 7
0
2
4
6
mRNAnormalizedby
RPLPO
CYP1A1
0
1
2
3
UGT1A1
0
2
4
6
NTCP
0
2
4
6
8
BSEP
Patented process
0,0
0,5
1,0
1,5
2,0
2,5
Day 7 Day 14 Day 21
µgalbumin/100ngofDNA
2D BIOMIMESYS®Hepatocyte
*** ***
***
BIOMIMESYS®Liver
-5%
15%
35%
55%
75%
95%
115%
135%
0,01 0,1 1 10 100
Viability
Chlorpromazine (µM)
BIOMIMESYS®Hepatocyte - Chronic
BIOMIMESYS®Hepatocyte - Acute
BIOMIMESYS®Liver - Chronic
BIOMIMESYS®Liver - Acute
BIOMIMESYS®Liver