CA-HPV-10 (ATCC® CRL-2220)

Organism: Homo sapiens, human  /  Cell Type: human papillomavirus 18 (HPV-18) transfected  /  Tissue: prostate  /  Disease: adenocarcinoma

Organism Homo sapiens, human
Tissue prostate
Cell Type human papillomavirus 18 (HPV-18) transfected
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 [Cells contain human papilloma viral DNA sequences]
Disease adenocarcinoma
Age 63 years
Gender male
Ethnicity Caucasian
Storage Conditions liquid nitrogen vapor phase
Karyotype aneuploid; mean chromosome number at passage 26 was 72; 10% of the cells retain the double minutes seen in the source tumor
Derivation
CA-HPV-10 was derived from cells from a prostatic adenocarcinoma of Gleason Grade 4/4. The cells were transformed by transfection with HPV18 DNA. Incorporation of HPV18 DNA was confirmed by polymerase chain reaction. Specific amplification of a 160-base pair fragment of the HPV18 E6 transforming region was noted. Immunocytochemical analysis showed expression of keratins 5 and 8 and also the early region 6 (E6) oncoprotein of HPV.
Clinical Data
63 years
Caucasian
male
Antigen Expression
kallikrein 3, KLK3 (prostate specific antigen, PSA); Homo sapiens
Tumorigenic No
Effects
No, The cells were not tumorigenic in immunosuppressed mice, but did form colonies in semisolid medium.
Comments

CA-HPV-10 was derived from cells from a prostatic adenocarcinoma of Gleason Grade 4/4. The cells were transformed by transfection with HPV18 DNA. Incorporation of HPV18 DNA was confirmed by polymerase chain reaction.

Immunocytochemical analysis showed expression of keratins 5 and 8 and also the early region 6 (E6) oncoprotein of HPV.

Complete Growth Medium The base medium for this cell line is provided by Invitrogen (GIBCO) as part of a kit: Keratinocyte Serum Free Medium (K-SFM), Kit Catalog Number 17005-042. This kit is supplied with each of the two additives required to grow this cell line (bovine pituitary extract (BPE) and human recombinant epidermal growth factor (EGF). To make the complete growth medium, you will need to add the following components to the base medium:
  • 0.05 mg/ml BPE - provided with the K-SFM kit
  • 5 ng/ml EGF - provided with the K-SFM kit. NOTE: Do not filter complete medium.
  • 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 0.1% Soybean Trypsin Inhibitor and aspirate cells by gently pipetting.
    5. To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. 
    6. Discard supernatant and resuspend cells in fresh growth medium.  Add appropriate aliquots of cell suspension to new culture vessels. 
    7. Incubate cultures at 37°C

    Subculture Ratio: 1:3 to 1:4
    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: Culture medium, 85%; fetal bovine serum, 10%; DMSO, 5%
    Storage temperature: liquid nitrogen vapor phase
    Culture Conditions
    Temperature: 37°C
    Name of Depositor DM Peehl
    References

    Weijerman PC, et al. Lipofection-mediated immortalization of human prostatic epithelial cells of normal and malignant origin using human papillomavirus type 18 DNA. Cancer Res. 54: 5579-5583, 1994. PubMed: 7923200

    Basic Documentation
    References

    Weijerman PC, et al. Lipofection-mediated immortalization of human prostatic epithelial cells of normal and malignant origin using human papillomavirus type 18 DNA. Cancer Res. 54: 5579-5583, 1994. PubMed: 7923200