Culture Collections

Hybridoma Detail

Important Advice: Resuscitation & Establishment of Hybridoma Cultures

ECACC Hybridoma Collection: W6/1

Supplied by: European Collection of Authenticated Cell Cultures (ECACC)
Culture Type: Cell line, hybridoma
Collection: ECACC Hybridoma Collection
Catalogue No.: 13082201
Cell Line Name: W6/1
Other Collection No.: ATCC N/A
Reactivity: Human 'A' erythrocytes
Antibody Isotype: IgM
Fusion Species: Mouse x Mouse Hybridoma
Myeloma: P3-NS1/1-Ag4.1
Immunological Donor: BALB/c mouse spleen
Immunogen: Human tonsil membrane
Transfected DNA: Not Applicable
Cell Line Description: The W6/1 hybridoma cell line secretes monoclonal IgM antibodies reactive with a polymorphic determinant of human erythrocytes which correlated with blood group A. Application: human erythrocyte marker.
Citation Guidance: If use of this culture results in a scientific publication, it should be cited in the publication as: W6/1 (ECACC 13082201)
Cell Type: Hybridoma
Morphology: Spherical
Growth Mode: Suspension
Sub Culture Routine: Seed at 5 x 104cells/ml. Subculture when cells reach 5 x 105-1 x 106cells/ml (usually after 3-4 days), 5% CO2, 37°C. Start in 20% FBS when recovering from frozen.
When recovering hybridoma cultures from frozen it is not unusual for growth to be slower than expected initially and there may be an observed decrease in viability. Establishment of an actively proliferating culture may take up to 2 weeks.
On resuscitation 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 prewarmed growth media+. Remove a sample for counting. Centrifuge at 100 x g for 2-3 minutes to pellet cells and seed at a relatively high density of 5-7 x 10⁵ cells/ml. Place culture flask flat and observe regularly until viable proliferating cells are seen.
Often hybridoma cultures may benefit from being resuspended with media supplemented with 20% FBS in the early critical stage of culture establishment immediately post resuscitation. Once the culture is established in culture the FBS can be reduced to 10%.
Culture Medium: RPMI 1640 + 2mM Glutamine + 10% Foetal Bovine Serum (FBS)
Depositor: Prof. A Neil Barclay, Cellular Immunology Unit, Sir William Dunn School of Pathology, University of Oxford
Originator: No
Country: UK
References: Barnstable CJ, Bodmer WF, Brown G, Galfre G, Milstein C, Williams AF, Ziegler A. 1978 Production of monoclonal antibodies to group A erythrocytes, HLA and other human cell surface antigens-new tools for genetic analysis. Cell. 14(1):9-20. PMID: 667938.
Patent: None Specified by Depositor
Release Conditions: Yes
Hazard Group (ACDP): 1
Biosafety: Unless specified otherwise, at the European Collection of Authenticated Cell Cultures (ECACC) we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines. ACDP = 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.

Hyperlinks to MSDS documents:
Frozen cell cultures Material Safety Data Sheet
Growing cell cultures Material Safety Data Sheet
Nucleic acids derived from cell cultures Material Safety Data Sheet

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.

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