Detachment factors for enhanced carrier to carrier transfer of CHO cell lines on macroporous microcarriers.
In this publication different detachment factors were tested for enhancing carrier to carrier transfer for scale-up of macroporous microcarrier based bioprocesses. Two Chinese hamster ovary cell lines, CHO-K1 and a genetically engineered CHO-K1 derived cell line (CHO-MPS), producing recombinant human Arylsulfatase B, were examined. The cells were grown on Cytoline 1microcarriers (Amersham Biosciences, Uppsala, Sweden) in protein-free and chemically defined medium respectively. Fully colonised microcarriers were used at passage ratios of approximately 1:10 for carrier to carrier transfer experiments. To accelerate the colonisation of the non-colonised, freshly added microcarriers the detachment reagents trypsin, papain, Accutasetrade mark (PAA, Linz, Austria), heparin and dextransulphate were used. Both cell lines showed good results with trypsin, Accutase and dextransulphate (Amersham Biosciences, Uppsala, Sweden), while papain failed to enhance carrier to carrier transfer in comparison to the non-treated reference. The maximum growth rate of cells on microcarriers with 2% dextransulphate in the medium was 0.25 +/- 0.02d(-1) and 0.27 +/- 0.03d(-1) for the CHO-MPS and CHO-K1, respectively. TheCHO-K1 grew best after detachment with trypsin (mu = 0.36 +/- 0.03d(-1)). This indicates, that one of the key parameters for carrier to carrier transfer is the uniform distribution of cells on the individual carriers during the initial phase. When this distribution can be improved, growth rate increases, resulting in a faster and more stable process.
Landauer
K
K
Institute of applied Microbiology, Muthgasse 18/Haus B, Wien, 1190, Austria.
Dürrschmid
M
M
Klug
H
H
Wiederkum
S
S
Blüml
G
G
Doblhoff-Dier
O
O