Description: Sp2/0-Ag14 cell line (ECACC catalogue no. 85072401) adapted to grow in the chemically defined protein- and peptide-free medium TurboDoma (Cell Culture Technologies). Suitable for use as a fusion parent for serum-free generation of hybridomas as well as a host for the production of recombinant proteins.

Also Known As:

Species: Mouse

Tissue: Unknown

Growth Properties: Suspension

Morphology: Lymphoblast

Growth Medium: TurboDoma medium (Cell Culture Technologies http://www.cellculture.com/) + 4mM Glutamine. When freezing cells down use TurboDoma medium (50:50 conditioned medium:fresh medium) + 4mM Glutamine + 10% DMSO.

Subculturing Procedure: Viability may be poor on resuscitation and may initially decrease further. Full recovery may take up to 2 weeks. A centrifugation step to remove the cryoprotectant is essential. Rapidly thaw the frozen ampoule in a water bath at 37°C for 1-2 minutes. Transfer the contents to a centrifuge tube and slowly add 5-10ml of pre-warmed growth media. Remove a sample for counting. Centrifuge at 100g for 2-3 minutes to pellet cells and seed at a relatively high density of 5-7 x 100,000 cells/ml. Leave culture flask upright and observe regularly until viable proliferating cells are seen. Once established use a split ratio of 1:2 approximately every 4 to 5 days; 5% CO2; 37°C.

Release Conditions: No

Depositor: Joint deposit from: Dr. Ferruccio Messi of Cell Culture Technologies Inc., Via al Chioso 14, 6929 Gravesano, Switzerland, Rene Fischer of the Laboratory for Organic Chemistry, ETH Zurich, 8093 Zurich, Switzerland, and Ing. Claudio Strebel of CePower Inc., Einsiedlerstr. 29, 8820 Waedenswil, Switzerland. Supported by the Swiss Foundation FFVFF

Originator: Yes

References: Shulman M, Wilde CD, Köhler G 1978 A better cell line for making hybridomas secreting specific antibodies. Nature. 276(5685):269-70 PMID: 714156. Ozato K, Sachs DH 1981 Monoclonal antibodies to mouse MHC antigens. III. Hybridoma antibodies reacting to antigens of the H-2b haplotype reveal genetic control of isotype expression. J Immunol. 126(1):317-21 PMID: 6935293.

Hyperlink to ECACC Cell Line Data Sheet: 03092501