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Sp2/0-Ag14 (serum-free)

Sp2/0-Ag14 (serum-free)

Catalogue No.


Cell Line Name

Sp2/0-Ag14 (serum-free)

Cell Line 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 20µg/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.


Tissue of Origin



Fusion partner


None Stated

Culture Conditions

Cell Type


Subculture Routine

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 10⁵ 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.

Culture 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)

Growth Mode


Additional Info


Dr. E. Penn, Health Protection Agency Culture Collections, Health Protection Agency, Porton Down, Salisbury, SP4 0JG, UK

Country of Origin

United Kingdom

Hazard Group (ACDP)

Hazard Group (ACDP) 2



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.

Available Formats

  • Frozen
  • DNA-5µg (100ng/µl)

If use of this culture results in a scientific publication, it should be cited in the publication as: Sp2/0-Ag14 (serum-free) (ECACC 08060101).

Unless specified otherwise, at ECACC we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines (Advisory Committee on Dangerous Pathogens) (UK). All cell cultures have the potential to carry as yet unidentified adventitious agents. It is the responsibility of the end user to ensure that their facilities comply with biosafety regulations for their own country. ACDP Guidance: Biological agents: Managing the risks in laboratories and healthcare premises.

The Culture Collections represent deposits of cultures from world-wide sources. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. References where quoted are mainly attributed to the establishment of the cell culture and not for any specific property of the cell line, therefore further references should be obtained regarding cell culture characteristics. Passage numbers where given act only as a guide and Culture Collections does not guarantee the passage number stated will be the passage number received by the customer.

Cultures supplied by Culture Collections are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.