3D culture scaffold for hepatocytes

3D culture scaffold for hepatocytes

BIOMIMESYS® Liver, a 3D cell culture model for maintaining and promoting hepatocytes functions for metabolism and toxicity studies

 

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. The SEM image shows the biomimetic structure of the BIOMEMISYS® Liver hydroscaffold with collagen chains.
 
A factor which greatly determines cell activity and viability is the extracellular matrix (ECM). Liver cells are organized and maintained by a complex extracellular microarchitecture. ECM compounds play a tremendous role in liver function regulation. Collagens, glycoproteins and glycosaminoglycans (GAG) constitute the majority of the ECM liver; also play important roles in cell physiology.

 

 
Biomimesys-liver-hepatocyte
BIOMIMESYS®Liver (on the right) and ECM from decellularized liver (on the left)
 
Hyaluronic acid (HA) has significant roles in the physical properties of liver tissue. HA sequesters water to maintain hydration of the ECM and also provides lubrication to enable cell motility and migration. HA is important during new liver tissue formation to promote activity of proliferating cells. These properties and its ability to avoid immune reactions upon transplantation make HA a suitable substrate for 3D cell culture and use for in vivo treatments.
 
Physicochemical features

The mechanical properties of BIOMIMESYS® Liver hydroscaffold display a Young's modulus of 0.6 kPa, which is in the same range of Liver ECM. The hydroscaffold is porous with pores size ranging from 60 µm to 130 µm diameter.
 
 
Spheroid: Viability&Growth: Increase Culture Time

Primary human hepatocytes and hepatocarcinoma cell line, as HepG2, remain viable for longer time in culture compared to 2D. Furthermore, primary human hepatocytes keep their metabolic activity for longer time in culture compared to 2D, even in sandwich culture.
 
Biomimesys-liver-HepG2
 
Maintains hepatocytes functions: Biliary Canaliculi Formation – HepG2

Biomimesys-liver-hepatocyte
Formation of canaliculi network with Actin and MRP2 colocalization
 
Maintains metabolic activities: Cryopreserved human hepatocytes

20 to 30 times more albumin is secreted in BIOMIMESYS®Liver than in 2D.
Basal CYP activity was analysed after 7 days of culture in 2D or in 3D.
 
 
Basal CYP activities are higher using BIOMIMESYS® Liver compared to 2D culture in sandwich.