Z Gastroenterol 2012; 50 - P2_05
DOI: 10.1055/s-0031-1295802

Characterization of liver specific functions of upcyte® hepatocytes grown in 3D and co-cultured with upcyte® endothelial cells

C Dähn 1, N Hewitt 2, D Maltman 3, G Talas 3, S Przyborski 3, S Heinz 2, A Nörenberg 2, K Scheller 2, J Braspenning 2
  • 1., .
  • 2Medicyte GmbH, Heidelberg
  • 3reinnervate, Sedgefield, UNITED KINGDOM


Major disadvantages of using human primary hepatocytes are their lack of proliferation and dedifferentiation in culture. Therefore, proliferating hepatocytes in combination with sophisticated 3D-culture systems capable of sustaining long-term functional cultures are highly desirable.Medicyte has developed a new technology that drives differentiated primary cells into proliferation in vitro, without uncontrolled cell growth or loss of phenotype. Upcyte® technology is based on targeted genetic modification of human primary cells allowing multiple passaging and the generation of billions of quasi primary cells.

Here, we present initial studies of the development of a 3D-cell culture model based on alvetex® scaffolds, containing upcyte® hepatocytes and microvascular endothelial cells (mvEC) in co-culture. Alvetex® scaffolds were kindly donated by Reinnervate Ltd, UK. Alvetex® is a highly porous, cross-linked polystyrene scaffold, which is sectioned into 200µm thick membranes. We aim to generate a more predictive in vitro model with improved liver specific functions compared to that seen in 2D-cell cultures.


Basal CYP3A4 activities of mono-cultures of upcyte® hepatocytes on alvetex® scaffolds were increased by up to 7-fold compared to 2D-cultures. Moreover, co-cultures with mvEC on alvetex® scaffolds resulted in 50-fold higher basal CYP3A4 activities than 2D cultures. Likewise, CYP2B6 activity was increased by 27-fold when upcyte® hepatocytes alone were cultured on alvetex® scaffolds compared to 2D cultures.


Upcyte® hepatocytes cultured on alvetex® scaffolds and in combination with upcyte® mvEC, showed improved CYP metabolism, indicating a more differentiated phenotype in 3D- compared to a 2D environment. These initial studies support the use of upcyte® hepatocytes in combination with non-parenchymal liver cells for further development of 3D culture systems which more closely resemble the in vivo structure of the liver.