human bladder epithelial cell line 5637  (ATCC)

Journal: BMC Urology

Article Title: Advances in experimental bladder models: bridging the gap between in vitro and in vivo approaches for investigating urinary tract infections

doi: 10.1186/s12894-024-01590-w

Figure Lengend Snippet: Comparison of different organoids used as bladder models

Article Snippet: • 5637 human bladder epithelial carcinoma cell line (procured from ATCC HTB-9TM) • Human primary bladder epithelial cells • Human Bladder Micro-vascular Endothelial cells (HMVEC-Bd) , UPEC strain CFT073 , To develop and characterize a bladder-chip model that mimics bladder structure by co-culturing human bladder epithelial cells with bladder microvascular endothelial cells, using a device that exposes epithelial cells to urine and endothelial cells to culture media , Neutrophils rapidly recruited from the vascular channel to infection sites formed swarms and extracellular traps, but this did not prevent the formation of intracellular bacterial communities (IBCs). Antibiotics showed delayed effectiveness in eliminating bacteria within IBCs, and in some cases, bacteria were not eradicated at all. During recovery periods, bacteria rapidly proliferated in IBCs, leading to new infection sites through bacterial shedding and host cell exfoliation. , [ ] .

Techniques: Comparison, Mutagenesis, Cell Culture, Expressing, Bacteria, Infection

Journal: bioRxiv

Article Title: Dynamic persistence of intracellular bacterial communities of uropathogenic Escherichia coli in a human bladder-chip model of urinary tract infections

doi: 10.1101/2021.01.03.424836

Figure Lengend Snippet: (A) Schematic of the human bladder-chip with co-culture of the HTB9 human bladder epithelial cell line (epithelium, top) and primary human bladder microvascular endothelial cells (endothelial, bottom) on either side of the stretchable and porous membrane. Pooled human urine diluted in PBS and endothelial cell medium were perfused in the apical and vascular channels respectively to mimic bladder physiology. A negative pressure in the ‘vacuum’ channels (magenta) on either side of the main channel was applied to stretch the porous membrane to mimic stretching of the bladder. ( B , C ) Immunofluorescence staining of confluent epithelial and endothelial cell monolayers (anti-EpCAM (magenta) and anti-CK7 (yellow) for the epithelial cells and anti-PECAM-1 (green) for the endothelial cells) in an uninfected control chip. Some endothelial cells also stained positive for CK7. Cell nuclei were labeled with DAPI (azure). ( D ) Schematic of the reconstitution of the bladder filling and voiding cycle via stretching of the membrane with a duty cycle of 6 hours. The cycle consisted of a linear increase in strain through stretching of the membrane ( filling bladder , 0 to 2 hours), maintenance of the membrane under stretch ( filled bladder , 2 to 4 hours), a quick relaxation of applied strain over 2 minutes ( voiding bladder , 4:02 hours) and maintenance without applied strain ( voided bladder , 4:02 hours to 6hours). ( E ) An overview of the timeline of the experimental protocol including infection, addition of neutrophils via the vascular channel, and two cycles of antibiotic treatment interspersed by two bacterial growth cycles. The consecutive bladder duty cycles are indicated.

Article Snippet: The HTB9 human bladder epithelial carcinoma cell line (procured from ATCC, 5637) was cultured in RPMI 1640 medium supplemented with 10% Fetal Bovine Serum (FBS) as recommended by the supplier.

Techniques: Co-Culture Assay, Membrane, Immunofluorescence, Staining, Control, Labeling, Infection