VK2/E6E7

The VK2/E6E7 cell line is a human vaginal epithelial cell line that was immortalized using the E6 and E7 oncogenes from human papillomavirus (HPV) type 16. This immortalization method extends the replicative lifespan of the cells while preserving many of the functional and morphological characteristics of normal vaginal epithelial cells. VK2/E6E7 cells are part of a panel of HPV16/E6E7-immortalized cell lines that also include ectocervical (Ect1/E6E7) and endocervical (End1/E6E7) epithelial cells, providing a comprehensive in vitro model for studying the biology of the female lower genital tract.

VK2/E6E7 cells exhibit a keratinocyte-like morphology and form loosely organized colonies in culture, distinct from the more stratified appearance of ectocervical cells. They express several epithelial differentiation markers, including cytokeratins CK8, CK18, CK19, and involucrin. Interestingly, they also express CK10 and CK13, markers associated with suprabasal differentiation in stratified squamous epithelium, suggesting that they retain key characteristics of normal vaginal epithelial cells. Additionally, VK2/E6E7 cells weakly express the secretory component (SC) of the polymeric immunoglobulin receptor, which is typically absent in vaginal tissue but is upregulated in this cell line. These features make VK2/E6E7 a valuable tool for studying vaginal epithelial function, host-pathogen interactions, and mucosal immunology, as well as for testing pharmacological agents intended for intravaginal application.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.