Mouse-human hybridoma X87X ceils were cultivated using a novel perfusion culture apparatus provided with three-settling zones to separate the cells from the culture medium by gravitational settling. The maximum viable cell density in a serum-free culture medium attained 3.0 x 10 v cells/ml, when the
High cell density perfusion culture of mouse-human hybridomas
β Scribed by Yoshiharu Takazawa; Michiyuki Tokashiki
- Publisher
- Springer
- Year
- 1989
- Tongue
- English
- Weight
- 409 KB
- Volume
- 32
- Category
- Article
- ISSN
- 1432-0614
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β¦ Synopsis
Five mouse-human hybridomas, H2, H3, V1, V2 and V6 cells secreting anti-virus human monoclonal immunoglobulin G (IgG) were cultured in serum-free media at high density in a settling perfusion culture vessel with an inner cell sedimentation zone. The H2, H3 and V6 cells reached a density of 107 cells/ml in 0.5% (w/v) BSA-ITES-eRDF (see Materials and methods). The H2 cells reached only 6.8 x 106 cells/ml in the absence of bovine serum albumin (BSA), but the addition of 0.02% (w/v) Pluronic F68 increased the maximum cell density to 1.1 Γ 107 cells/ml, which was the same level as in BSA including medium. On the other hand, Pluronic F68 showed no stimulative effect on the growth of H2 cells in static culture. Pluronic F68 also increased the maximum cell density of V2 cells from 4.6 x 10 6 cells/ml to 6.9 Γ 106 cells/ml even in the presence of 0.5% (w/v) BSA.
π SIMILAR VOLUMES
Two mouse myeloma cell lines which were transfected with chimeric mouse variable-human constant immunoglobulin heavy and light chain genes have been cultured at high cell density in a settling perfusion culture vessel to produce chimeric antibody specific for human common acute lymphocytic leukemia
We have developed a high cell density and high product concentration culture system recycling high molecular weight components. The production of monoclonal antibodies in high concentration was performed by this culture system with mouse human hybridoma H2 and V6 cells in serum-free defined media.
The influence of centrifugal force on the growth of cells was examined by exposing the cells of the mouse-human hybridoma X87 line to centrifugal force (100-500 G) for ten minutes twice a day and comparing the static culture with that of unexposed cells. In this experiment, both cell proliferation a