Description: Sp2/0-Ag14 cell line (ECACC catalogue no. 85072401) adapted to grow in animal-component free, protein-free, chemically defined, serum-free medium (EX-CELL® Sp2/0 Serum-Free Medium for Sp2/0 Cells Chemically Defined, Sigma cat no 14660C). Sp2/0-Ag14 is a non-Ig-secreting or synthesising line derived from a cell line created by fusing a BALB/c mouse spleen cell and the mouse myeloma P3X63Ag8. Resistant to 8-azaguanine at 20ug/ml and does not survive in HAT containing media. Can be used as a fusion partner for generating hybridomas. This cell line was previously known as SP2/0-Ag14 (AC-Free) however the name has been changed to Sp2/0-Ag14 (serum-free) to make the serum free status of this culture more obvious to users.

Also Known As: SP2/0-Ag14 (AC-Free)

Species: Mouse

Tissue: Unknown

Growth Properties: Suspension

Morphology: Lymphoblast

Growth Medium: EX-CELL Sp2/0 Serum-Free Medium (Sigma cat no. 14660C) + 8mM Glutamine. When freezing cells down use 50:50 fresh culture medium:conditioned medium plus 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; 10% CO2; 37°C.

Release Conditions: No

Depositor:

Originator: No

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: 08060101