title: Comparison of Chlamydia trachomatis serovar L2 growth in polarized genital epithelial cells grown in three-dimensional culture with non-polarized cells.
abstract: A common model for studying Chlamydia trachomatis and growing chlamydial stocks uses Lymphogranuloma venereum serovar L2 and non-polarized HeLa cells. However, recent publications indicate that the growth rate and progeny yields can vary considerably for a particular strain depending on the cell line/type used, and seem to be partially related to cell tropism. In the present study, the growth of invasive serovar L2 was compared in endometrial HEC-1B and endocervical HeLa cells polarized on collagen-coated microcarrier beads, as well as in HeLa cells grown in tissue culture flasks. Microscopy analysis revealed no difference in chlamydial attachment/entry patterns or in inclusion development throughout the developmental cycle between cell lines. Very comparable growth curves in both cell lines were also found using real-time PCR analysis, with increases in chlamydial DNA content of 400-500-fold between 2 and 36 h post-inoculation. Similar progeny yields with comparable infectivity were recovered from HEC-1B and HeLa cell bead cultures, and no difference in chlamydial growth was found in polarized vs. non-polarized HeLa cells. In conclusion, unlike other C. trachomatis strains such as urogenital serovar E, invasive serovar L2 grows equally well in physiologically different endometrial and endocervical environments, regardless of the host cell polarization state.
authors.affiliation: Department of Microbiology, James H Quillen College of Medicine, East Tennessee State University, Johnson City, TN, USA.
authors.forename: Sophie
authors.initials: S
authors.lastname: Dessus-Babus
authors.forename: Cheryl G
authors.initials: CG
authors.lastname: Moore
authors.forename: Judy D
authors.initials: JD
authors.lastname: Whittimore
authors.forename: Priscilla B
authors.initials: PB
authors.lastname: Wyrick
mesh_terms: D002692:Chlamydia trachomatis / Q000254:growth & development* / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000032:analysis; D004717:Endometrium / Q000382:microbiology* / Q000648:ultrastructure; D005260:Female; D006367:HeLa Cells; D006801:Humans; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D008863:Microspheres; D016133:Polymerase Chain Reaction
chemical_list: D004269:DNA, Bacterial