Rotary cell culture system (RCCS): a new method for cultivating hepatocytes on microcarriers

The Rotary Cell Culture System (RCCS) is a new technology for growing ancho rage dependent or suspension cells in the laboratory The RCCS is a horizont ally rotated bubble free disposable culture vessel with diffusion gas excha nge. The system provides a reproducible, complex 3D in vitro culture system with large cell masses. During cell growing the rotation speed can be adju sted to compensate for increased sedimentation rates. The unique environmen t of low shear forces, high mass transfer and microgravity, provides very g ood cultivating conditions for many cell types, cell aggregates or tissue p articles in a standard tissue culture laboratory. The system enables to culture HepG2 cells on Cytodex 3 microcarriers (mcs) to high densities. We inoculated 2 x 10(5)/ml HepG2 cells and 200 mg Cytode x 3 mcs in 50 ml Williams E medium (incl. 10% FCS) allowing them to attach to the mcs in the rotating vessel (rotation rate 14-20 rpm). HepGP cells re adily attached to the mcs while the vessel was rotating. Attachment of HepG 2 to the mcs was about 50% after 24 hrs and 100% within 48 hrs. After 72 hr s of rotary culturing small aggregates of Hep G2 on mcs were built. HepG2 c ells and the aggregates rotated with the vessel and did not settle within t he vessel or collide with the wall of the vessel. We conclude that this new RCCS is an excellent technology for culturing Hep GP cells on Cytodex 3 mcs. The system is easy to handle and enables to cult ure anchorage dependent cells to high densities in a short period.