Biomedical Engineering Reference
In-Depth Information
56.
Int J Artif Organs
. 1999 Dec;
22(12):816-22. Rotary cell culture
system (RCCS): a new method
for cultivating hepatocytes on
microcarriers. Mitteregger R,
Vogt G, Rossmanith E,
Falkenhagen D. Christian
Doppler Laboratory for Specific
Adsorption Technologies in
Medicine, Krems, Austria.
The Rotary Cell Culture System (RCCS) is a new
technology for growing anchorage-dependent or
suspension cells in the laboratory. The RCCS is a
horizontally rotated, bubble-free disposable culture
vessel with diffusion gas exchange. The system
provides a reproducible, complex 3D in vitro culture
system with large cell masses. During cell growing,
the rotation speed can be adjusted to compensate for
increased sedimentation rates. The unique
environment of low shear forces, high mass transfer,
and microgravity provides very good cultivating
conditions for many cell types, cell aggregates, or
tissue particles in a standard tissue culture
laboratory. The system enables culturing of HepG2
cells on Cytodex 3 microcarriers (mcs) to high
densities. We inoculated 2 × 10(5)/mL HepG2 cells
and 200 mg Cytodex 3 mcs in 50 mL Williams E
medium (incl. 10% fetal calf serum (FCS), allowing
them to attach to the mcs in the rotating vessel
(rotation rate 14-20 rpm). HepG2 cells readily
attached to the mcs while the vessel was rotating.
Attachment of HepG2 to the mcs was about 50%
after 24 h and 100% within 48 h. After 72 h of rotary
culturing, small aggregates of Hep G2 on mcs were
built. HepG2 cells and the aggregates rotated with
the vessel and did not settle within the vessel or
collide with the wall of the vessel. We conclude that
this new RCCS is an excellent technology for
culturing HepG2 cells on Cytodex 3 mcs. The system
is easy to handle and enables culturing of anchorage-
dependent cells to high densities in a short period.
