Human Vaginal Epithelial Cell (HVEC) Line

Tool Details

*FOR RESEARCH USE ONLY (for other uses, please contact the licensing team)

• Name: Human Vaginal Epithelial Cell (HVEC) Line
• Alternate name: HVEC E6/E7/TERT cells (V428)
• Organism: Human
• Tissue: Vaginal Epithelium
• Disease: S. aureus infectioni
• Growth properties: Adherent
• Model: Immortalised Line
• Conditional: No
• Description: Immortalized HVEC line derived from primary normal human epithelial cells isolated from premenopausal vaginal hysterectomy tissue, obtained from a healthy female donor. This cell line maintains phenotype with time in culture and exhibits proinflammatory responses, and is useful for researchers studying cancer/infection biology and pathogenesis.
• Production details: Primary normal human epithelial cells were isolated from premenopausal vaginal hysterectomy tissue, obtained from a woman who did not have cancer. Cells were grown in keratinocyte serum-free medium (KSFM) (Gibco) on plastic and passaged with a 1:4 split using trypsin-EDTA. Early-passage cells were doubly transduced with retroviruses expressing human papillomavirus 16 (HPV-16) E6/E7 and the reverse transcriptase component of telomerase, hTERT, and selected with 50 μg of the antibiotic G418 per ml (Sigma). The E6/E7/TERT cells (V428) had high levels of telomerase and became immortal without a crisis, whereas normal un-transduced cells senesced around passage 9.
• Biosafety level: 2

Target Details

• Target: Used to assess affects of toxins/pathogen on vaginal epithelium

Handling

• Format: Frozen
• Growth medium: Keratinocyte serum-free medium (KSFM) (GibcoTM), supplemented with 25 ug/ml bovine pituitary extract (BPE) and 0.16 ng/ml epidermal growth factor (EGF) and a 2% final volume of penicillin-streptomycin-amphotericin B. Do not keep media for more than 1 month past adding supplements. Do not freeze and thaw medium.
• Temperature: 37°C
• Unit size: 1x10^6 cells / vial
• Shipping conditions: Dry ice
• Subculture routine: Split 1:4 every 4-5 days at 70-90% confluency. Aspirate medium and wash plate with (1X) PBS (without calcium or magnesium). Add 4 ml (0.05%) Trypsin/EDTA and incubate at 37°C for 5-10 minutes. Add 4 ml PBS-A containing 2% FBS to inactivate the trypsin. Transfer cells to a sterile 15ml tube and centrifuge at 1200 rpm for 5min. Carefully aspirate supernatant taking care not to disturb the pellet and resuspend in K-SFM. Change media every other day. The cells will not do well if they are left over the weekend without a media change or if they are left to grow to greater than 95% confluency. Please also see detailed protocol within the Product Datasheet in the Documentation section below.

References

• Peterson et al. 2005. ASM Journals. Infect Immun. 2005 Apr:73(4):2164–2174. PMID:15784559

Documentation

• Product Datasheet