2H-11 (ATCC® CRL-2163)

Organism: Mus musculus, mouse  /  Cell Type: endothelial, SV40 transformed  /  Tissue: axillary lymph node/vascular epithelium  / 

Organism Mus musculus, mouse
Tissue axillary lymph node/vascular epithelium
Cell Type endothelial, SV40 transformed
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 [Cells contain polyomavirus DNA sequences]
Age adult
Gender Male
Strain C3H/HeJ
Storage Conditions liquid nitrogen vapor phase
Derivation
The 2H-11 cell line was derived from a solid tumor in nude mice injected with SVEC4-10EHR1 (see ATCC CRL-2161).
The line was cloned in 1992 by limiting dilution.
Clinical Data
male
Antigen Expression
H-2 K; VCAM
Tumorigenic Yes
Effects
Yes, the cells induce spindle tumors with some of the histopathologic characteristics of human Kaposi Sarcoma after a latency period of approximately 14 weeks
Comments

The cells are sensitive to lysis by activated macrophages as measured in the chromium release assay.

This clone retains the ability to differentiate on a synthetic basement-like membrane.

The cells express the cell surface major histocompatibility complex class I antigen, H-2 k, of the parental cell line, and express VCAM (vascular cell adhesion molecule).

The cells stain positively for SV40 T antigen.

Complete Growth Medium Dulbecco's modified Eagle's medium with 4.5 g/L glucose, 90%; heat-inactivated fetal bovine serum, 10%
Subculturing Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C

Subculture Ratio: 1:4 to 1:6
Medium Renewal: 2 to 3 times a week.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation
Culture medium, 95%; DMSO, 5%
Name of Depositor KA O'Connell
References

O'Connell KA, Edidin M. A mouse lymphoid endothelial cell line immortalized by simian virus 40 binds lymphocytes and retains functional characteristics of normal endothelial cells. J. Immunol. 144: 521-525, 1990. PubMed: 2153170

O'Connell KA, Rudmann AA. Cloned spindle and epithelioid cells from murine Kaposi's sarcoma-like tumors are of endothelial origin. J. Invest. Dermatol. 100: 742-745, 1993. PubMed: 8496612

O'Connell K, et al. Endothelial cells transformed by SV40 T antigen cause Kaposi's sarcomalike tumors in nude mice. Am. J. Pathol. 139: 743-749, 1991. PubMed: 1928299

Basic Documentation
References

O'Connell KA, Edidin M. A mouse lymphoid endothelial cell line immortalized by simian virus 40 binds lymphocytes and retains functional characteristics of normal endothelial cells. J. Immunol. 144: 521-525, 1990. PubMed: 2153170

O'Connell KA, Rudmann AA. Cloned spindle and epithelioid cells from murine Kaposi's sarcoma-like tumors are of endothelial origin. J. Invest. Dermatol. 100: 742-745, 1993. PubMed: 8496612

O'Connell K, et al. Endothelial cells transformed by SV40 T antigen cause Kaposi's sarcomalike tumors in nude mice. Am. J. Pathol. 139: 743-749, 1991. PubMed: 1928299