Penicillin streptomycin p s

HeLa (human cervical cancer cell line) were cultured in Dulbecco’s Modified Eagle Medium with phenol red, 4.5 g L⁻¹d-glucose, l-glutamine and pyruvate (DMEM, 1X, Gibco) supplemented with 10% FBS (Gibco) and 1% Penicillin/Streptomycin (P/S, Corning, 100X). Cells were maintained under humid conditions at 37 °C and 5% of CO₂. Cells were passaged after cleaning Dulbecco’s Phosphate Buffered Saline (DPBS, 1X, Gibco) with 0.25% Trypsin–EDTA (1X, Gibco) when the culture reached confluency.

Vero cells (monkey kidney epithelial cells, Cat. # CCL-81) were purchased from ATCC (Manassas, VA, USA). The cells were grown at 37 °C and 5% CO₂ in complete Dulbecco’s modified Eagle medium (DMEM) medium: DMEM (Corning) supplemented with 10% fetal bovine serum (FBS) (Gibco) and 1% penicillin-streptomycin (P/S; Corning, Glendale, Arizona, US). The CHIKV French La Reunion strain LR2006-OPY1 was a kind gift of The Connecticut Agricultural Experiment Station located in New Haven, CT, USA. The ONNV non-recombinant strain was provided by the World Reference Center for Emerging Viruses and Arboviruses (WRCEVA) at the University of Texas Medical Branch. Both viruses were propagated in Vero cells.

Pericytes were seeded at 8000 cells/mL into two 6‐well cell culture plate (Corning Inc., Corning, NY, USA) precoated with gelatin (Cell Biologics Inc.) in either Dulbecco's modified Eagle media (DMEM) with 4.5 g·L⁻¹ glucose, l‐glutamine, and sodium pyruvate (Corning Inc.) supplemented with +5% FBS (Corning Inc.) +1% penicillin/streptomycin (P/S) (Corning Inc.) or endothelial growth medium 2 (EGM‐2; Cell Biologics, Inc., Chicago, IL, USA) supplemented with the ECs medium supplement kit (Catalog M1168, Cell Biologics Inc., Chicago, IL, USA). Cells were seeded at 10 000 cells per well into a 24‐well cell culture plate (Corning) precoated with gelatin (Cell Biologics Inc.) in cell culture media. After 6 h for cells to attach, media was changed to include LDL (Sigma) or d‐glucose (Sigma). Plates were placed into an Incucyte Zoom instrument (EssenBioscience Inc., Ann Arbor, MI, USA) integrated with the incucyte zoom 2018A software (EssenBioscience Inc., Ann Arbor, MI, USA), and growth was monitored for 5 days to construct proliferation curves.

Three immortalized human fimbrial epithelial cell lines (FT282-V, FT282-cyclin E1 (CCNE1), FE25), one subline derived from the xenograft tumor of FE25 cells (FEXT2), and two genomically-proven HGSC cell lines (OVSAHO and KURAMOCHI) were used in this study. The relevant genotypes and phenotypes are shown in Table 1. We established FE25 cells by transduction using human papillomavirus (HPV) E6/E7 plus human telomerase reverse transcriptase (hTERT) [30 (link)]. FT282-CCNE1 and FT282-V cell lines, a kind gift from Dr. Ronny Drapkin, were transduced with TP53 p.R175H, hTERT plus CCNE1 or vector, respectively [23 (link)]. These cells were maintained in MCDB105/M199 medium (1:1, Merck, NJ, USA) supplemented with 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Waltham, MA, USA) and penicillin/streptomycin (P/S, Corning Inc., Corning, NY, USA). The HGSC cell lines, KURAMOCHI and OVSAHO were obtained from the JCRB cell bank, Japan. The cells were cultured in RPMI 1640 medium (Gibco-Thermo Fisher Scientific, MA, USA) supplemented with 10% (v/v) FBS, penicillin (50 U/mL) and streptomycin (50 μg/mL).