RCCSTM Bioreactors to study Hepatitis C infection in-vitro
Synthecon, Inc

In a recent study conducted by scientists at the University of Illinois, Chicago, Synthecon, Inc. Rotary Cell Culture SystemTM was used to construct a Huh 7 3-D cell culture model (human hepatoma-derived cell line) that was superior to conventional 2-D monolayers in terms of increased hepatocyte-specific gene expression and cellular polarity.

These 3D cultures were able to be infected by HCV, thus establishing that HCV entry into 3-D cultured Huh7 cells is not limited by polarity. According to Dr. Susan Uprichard, “These more in vivo-like Huh7 cell cultures should prove useful for understanding the interplay between HCV and the cellular tight junction proteins that mediate infectious HCV entry into cells as well as how HCV affects other aspects of hepatocyte physiology”.

This study has demonstrated that the conditions of suspension and free-fall in the Rotary Cell Culture SystemTM would allow hepatocytes to differentiate into 3-D aggregates which are structurally and functionally similar to their in-vivo counterparts; thereby representing a more physiologically relevant model to study HCV infection dynamics. Synthecon expects that this kind of a high- fidelity 3-D hepatocyte model would be very useful not only to viral hepatitis work, but also to various other fields of liver research that require such a model.

Reorganization of HCV receptor, cell adhesion and tight junction protein localization in 3-D Huh7 aggregates (above picture as title)

Fourteen days post seeding, Huh7 3-D aggregates and parallel Huh7 2-D confluent monolayers were stained with antibodies specific for SR-BI, CD81, CLDN1, CD26, β-Catenin, E-cadherin, ZO-1 or Occludin and visualized via confocal microscopy (630×, Zeiss LSM 510, Germany).

According to the authors, “This data demonstrates that the expression and distribution of cell adhesion and TJ proteins, including known HCV entry receptors, are enhanced and more polarized in 3-D Huh7 cultures than in 2-D monolayers”. (This is conclusion from picture) (below picture as explanation)