PZ-HPV-7 (ATCC® CRL-2221)

Organism: Homo sapiens, human  /  Cell Type: human papillomavirus 18 (HPV-18) transformed, epithelial  /  Tissue: prostate; epithelium  / 

Organism Homo sapiens, human
Tissue prostate; epithelium
Cell Type human papillomavirus 18 (HPV-18) transformed, epithelial
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2
[Cells contain human papilloma viral DNA sequences]
Age 70 years adult
Gender male
Ethnicity Caucasian
Storage Conditions liquid nitrogen vapor phase
Karyotype at low passages maintained the diploid karyotype of the normal parental cells but by passage 99 the karyotype had changed to near-triploid.
Derivation
PZ-HPV-7 was derived from epithelial cells cultured from normal tissue from the peripheral zone of the prostate.
Specific amplification of a 160-base pair fragment of the HPV18 E6 transforming region was noted.
Incorporation of HPV18 DNA was confirmed by polymerase chain reaction.
The cells were transformed by transfection with HPV18 DNA.
Clinical Data
70 years adult
Caucasian
male
Tumorigenic No
Effects
No, The cells were not tumorigenic in immunosuppressed mice, but did form colonies in semisolid medium.
Comments
Immunocytochemical analysis showed expression of keratins 5 and 8 and also the early region 6 (E6) oncoprotein of HPV.

The cells are negative for prostate specific antigen (PSA).

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. 

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. Add 2.0 to 3.0 mL 0.25% (w/v) Trypsin-0.53 mM EDTA solution for 2-3 minutes. Add 6.0 to 8.0 ml of 0.1% Soybean Trypsin Inhibitor and gently dislodge the cells by agitating or tapping the flask. 
    3. To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
    4. Discard supernatant and resuspend cells in fresh growth medium.  Add appropriate aliquots of cell suspension to new culture vessels. 
    5. Incubate cultures at 37°C

    Subculture Ratio: 1:3
    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, 85%; fetal bovine serum, 10%; DMSO, 5%
    Storage Temperature: Liquid nitrogen vapor temperature
    Culture Conditions
    Temperature: 37°C
    STR Profile
    Amelogenin: X,Y
    CSF1PO: 11,12
    D13S317: 12,14
    D16S539: 11,12
    D5S818: 9,13
    D7S820: 9
    THO1: 7,9
    TPOX: 8,11
    vWA: 17
    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