YPEN-1 (ATCC® CRL-2222)

Organism: Rattus norvegicus, rat  /  Cell Type: immortalized with adenovirus 12 - SV40 virus hybrid (Ad12-SV40)  /  Tissue: prostate, endothelium  /  Disease: normal

Organism Rattus norvegicus, rat
Tissue prostate, endothelium
Cell Type immortalized with adenovirus 12 - SV40 virus hybrid (Ad12-SV40)
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 Cells contain polyomavirus DNA sequences
Disease normal
Age 8 weeks
Gender male
Strain Copenhagen
Storage Conditions liquid nitrogen vapor phase
Derivation
The YPEN-1 cell line was originated in 1993 from prostate cells of 8 week old Copenhagen male rats.
Tumorigenic No
Effects
No, the cells were not tumorigenic in immunosuppressed mice, but did form colonies in semisolid medium.
Comments

Cells were cultured in Endothelium Isolation Medium and immortalized by infection with Adenovirus12 SV40 hybrid virus.

The cells stain positively for but are non-producers of SV40 T-antigen.

YPEN-1 cells demonstrate acetylated low density lipoprotein (Dil-Ac-LDL) uptake as an endothelial marker.

They exhibit positive staining for endothelin and for a monoclonal antibody to rat endothelium (MRC OX-43).

They express Integrin a6 1 and Integrin 3 on their plasma membrane, and demonstrate tube formation in Matrigel.

Complete Growth Medium Minimum essential medium (Eagle) with 2 mM L-glutamine and Earle's BSS adjusted to contain 1.5 g/L sodium bicarbonate, 0.1 mM non-essential amino acids, and 1.0 mM sodium pyruvate, supplemented with 0.03 mg/ml heparin, 95%; fetal bovine serum, 5%
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 which 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.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended
Medium Renewal: Every 2 to 3 days

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
Freeze medium: Complete growth medium, 95%; DMSO, 5%. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
Culture Conditions
Temperature: 37°C
Population Doubling Time 26 hrs
Name of Depositor K Yamakazi, K Pienta
Year of Origin 1993
References

Yamazaki K, et al. Establishment of immortalized Copenhagen rat prostate endothelial cell lines. In Vivo 9: 421-426, 1995. PubMed: 8900918

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm

Basic Documentation
References

Yamazaki K, et al. Establishment of immortalized Copenhagen rat prostate endothelial cell lines. In Vivo 9: 421-426, 1995. PubMed: 8900918

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm