New perfluorocarbon system for multilayer growth of anchorage-dependent mammalian cells

A novel tissue culture system has been developed that supports the multilayer growth of Hep G2 cells. The system consists of growing cells on collagen-coated perfluorocarbon substrata in the wells of a multi-well plate designed so that, even at very high densities, the oxygen in the cultures is replenished as rapidly as it is consumed. Hep G2 cells, which are typically contact inhibited, grow to form more than 10 layers of cells that continue to secrete albumin. Both multilayer growth and high rates of albumin depend on using a very enriched nutrient medium, compared to media usually used for monolayer culture of Hep G2 cells. The role played by increased oxygenation, enriched media, and the unique properties of the perfluorocarbon substrata for the 3-D growth of anchorage-dependent cells is discussed.