HuH-7 is a well differentiated hepatocyte derived cellular carcinoma cell line that was originally taken from a liver tumor in a 57-year-old Japanese male in 1982. The line was established by Nakabayshi, H. and Sato, J.
HuH-7 is an immortal cell line of epithelial-like tumorigenic cells. It usually grow in 2D monolayers.
To use as a hepatocyte culture model, HuH-7 was cultured in 3-D Rotating Wall Vessel (RWV) bioreactors engineered by NASA. This is a horizontally rotating cylindrical culture vessel that is meant to grow cells in 3D aggregates, without the shear and turbulence associated with conventional, stirred bioreactors. Cells cultured this way are grown much like the cells of a fetus in a mother’s uterus. They experience an environment with aspects of microgravity and “suspended animation”, where the cells are in constant free-fall. The fluid dynamics of the RWV cell culture allows cells to develop more naturally, in a 3D space, much like they would in vivo. This is in contrast to the 2D plastic substrates in which most cells are grown in culture (plates). Thus, RWV grown cells are more similar, in structure and function, to their in vivo counterparts, resulting in a more physiologically relevant system for the study of viruses.
The difference between RWV and conventional cell culture is remarkable, and completely altered gene expression and differentiation is often observed in RWV cultured cells. RWV promotes differentiation of the HCV receptive HuH-7 cells. The RWV grown HuH-7 cells are distinct from original HuH-7 cells both morphologically, and transcriptionally. In the RWV, these cells form bulky aggregates which are more polarized than the original cells. RWV cells also exhibit greater regulation of metabolic and hepatocyte-specific transcripts, as well as increased expression of proteins necessary for the formation of tight junctions and cell adhesion.