| 產(chǎn)品名稱 |
PZ-HPV-7 |
| 商品貨號 |
B165541 |
| 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]
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| 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.
- Remove and discard culture medium.
- 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.
- To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
- Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
- 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 |
| Deposited As |
Homo sapiens |
| 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
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