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UK Health Security Agency

8C9

8C9

Catalogue No.

99072805

Cell Line Name

8C9

Cell Line Description

The hybridoma line 8C9 was produced by immunizing a female BALB/c mouse i.p. with tissue from a human malignant fibrous histiocytoma once a week for 6 weeks. Mouse spleen cells were fused with myeloma NS-1 cells using PEG and HAT for selection . The 8C9 monoclonal antibody detects a unique epitope on the leukocyte differentiation antigen CD36 (platelet glycoprotein IV or III6) expressed by normal and neoplastic endothelial cells. Immunoreactivity with 8C9 was detected on capillary endothelial cells, adipocytes, monocytes, platelets and the human monocytoid cell line U-937 (ECACC catalogue no. 85011440). The antibody did not react with erythroid cells, T cells, B cells or HUVECs (human umbilical vein endothelial cells). Immunoblot analysis showed that 8C9 bound to a 97-kDa membrane protein expressed by U-937 cells treated with phorbol ester. Comparative immunohistochemical studies using 8C9 and the monoclonal antibody OKM demonstrated epitopic heterogeneity of the CD36 antigen by normal and neoplastic endothelial cells.

Characteristics

Myeloma

NS-1

Immunogen

Human malignant fibrous histiocytoma

Immunological Donor

BALB/c female mouse spleen

Antibody Isotype

IgM

Morphology

Spherical

Tissue of Origin

Hybrid

Culture Conditions

Cell Type

Hybridoma

Subculture Routine

Maintain cultures between 3-9 x 100,000 cells/ml; 5% CO₂; 37°C. 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 10µ 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% FBS

Growth Mode

Suspension

Additional Info

Depositor

Obtained from Riken Cell Bank

Country of Origin

Japan

Applications

Study of endothelial cells, CD36 expression analysis

Hazard Group (ACDP)

2

References

References

Acta Pathol Jpn 1992; 42:807

Available Formats

  • Frozen

If use of this culture results in a scientific publication, it should be cited in the publication as: 8C9 (ECACC 99072805)

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.