Immortalized Human Endocervical Epithelial Cells- HPV E6/E7 (A2EN)

The endocervical epithelium lines the endocervix in the female reproductive tract, separating sterile and microbe-rich regions from one another. Since the endocervix is a target for sexually transmitted organisms such as Chlamydia trachomatis and Neisseria gonorrhoeae, the endoecervical epithelial cells play an important role in both host defense and reproduction. The Immortalized Endocervical Epithelial Cells – HPV E6/E7 is unique in that it can be polarized to exhibit distinct apical and basolateral membrane domains and that it retained its ability to respond to hormone and microbial stimulus. A2EN cells serve as an excellent model to further characterization of pathogen-host interactions on sexually transmitted infections.

Immortalization Method: Serial passaging and transduction with retroviruses carrying Human papillomavirus E6/E7 (type 16) protein (pLXSN)

BioSafety Level: II

Species: Human

Source Organ: Endocervix

Growth Properties: Adherent

Morphology: Cobble-stone

Population Doubling: N/A

Seeding Density: Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.

Markers: MUC5B

Donor Age: N/A

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: The base medium for this cell line is Prigrow X.1 medium available at abm, Cat. No. TM010. To make the complete growth medium, add the entire content of the growth supplements to the basal media (no additional fetal bovine serum is required) and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.
Carbon dioxide (CO₂): 5%, Temperature: +37.0°C.

Quality Control: (1) Electron microscopy was performed to assess cell polarization, while fluorescent microscopy was performed to assess tight junction formation between cells. (2) Transepithelial electrical resistance was measured to measure cell monolayer functional integrity. (3) RT-PCR, Western blotting, and immunostaining were performed to compare immortalized cell characteristics to normal counterparts. (4) Expression levels of antimicrobial peptides, cytokines, and chemokines were measured to further assess immortalized cell characteristics. (5) qPCR was performed to analyze toll-like receptor protein expression between basolateral and apical components of cells, while qRT-PCR was performed to determine hormone receptor functionality.

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C