human bladder epithelial cell line 5637  (ATCC)

Journal: bioRxiv

Article Title: Unmasking Pathogen Traits for Chronic Colonization in Neurogenic Bladder Patients

doi: 10.1101/2025.08.14.669717

Figure Lengend Snippet: a, Schematic of LPS with the R1-type core oligosaccharide. Geometric symbols represent sugar subunits of the oligosaccharide portions of LPS. Abbreviations: Kdo—3-deoxy-d-manno-oct-2-ulosonic acid; Hep—glycero-d-manno-heptose; Glc—glucose; Gal—galactose. b, Premature stop codon in waaW from E. coli isolate BTF6T2 relative to isolate BTF6T1 from the same patient. Mutation visualized with Snipit . c, Silver stained LPS extracts. The LPS standard--is a commercially available product from E. coli serotype O55:B5 (1.25 µg of LPS standard was added). d, Adherence to and invasion of bladder epithelial cells (ATCC HBT9). UTI89 and its isogenic mutants waaW and waaL were tested. e, Macrophage internalization and survival of UTI89 and its isogenic waaW mutant. Internalized CFUs were counted 30 minutes after inoculation, and surviving CFUs were counted from within the macrophages 90 minutes after inoculation. For panels d and e , the horizontal line is the geometric mean, and the error bars depict the 95% CI. P values were calculated with non-parametric Kruskal–Wallis test with two-sided Dunn’s post hoc test for multiple comparisons.

Article Snippet: Bladder epithelial cells (ATCC 5637, HTB9) were maintained in RPMI 1640 Media (Gibco A1049101) with 10% heat inactivated fetal bovine serum (FBS), 100 IU/mL of penicillin and 100 μg/mL of streptomycin.

Techniques: Mutagenesis, Staining

Journal: bioRxiv

Article Title: Lysine Decarboxylation aids in UPEC intracellular survival in the early stages of urinary tract infection

doi: 10.1101/2025.04.25.650641

Figure Lengend Snippet: Graph shows RT-qPCR results of representative genes from each AR system, using RNA isolated from intracellular bacteria. For this work the bladder epithelial cell line 5637 (ATCC HTB-9) was seeded with wild-type UTI89 at an MOI=10. Following a 2h incubation to allow for pathogen internalization, extracellular bacteria were eliminated by incubating the urothelial cell line with gentamicin for another 2 hours. Gentamicin was then removed and cells were washed 3X with 1X PBS, followed by urothelial cell lysis and subsequent treatment for bacterial lysis and RNA extraction. Extracted RNA was DNAse-treated, reverse-transcribed, quantified and subjected to qPCR using TaqMan probes designed for adiA (purple), cadA (green), gadA (pale blue), gadC (light blue), sdaA (red), sdaC (royal blue), and speF (coral). Transcript abundance was normalized to the gyrB house keeping gene transcripts. Relative fold changes were determined by the ΔΔ C T method, relative to the bacterial inoculum corresponding transcripts. Graph shows analysis of 4 biological replicates. Error bars indicate mean and standard error of the mean at 95% CI.

Article Snippet: Cell culture infections were performed using 5637 human transitional bladder epithelial cells (ATCC HTB-9) originally derived from a 68-year old white male with Grade II bladder carcinoma.

Techniques: Quantitative RT-PCR, Isolation, Bacteria, Incubation, Lysis, RNA Extraction, Reverse Transcription