human female urine  (Golden West Biologicals)

Journal: eLife

Article Title: Dynamic persistence of UPEC intracellular bacterial communities in a human bladder-chip model of urinary tract infection

doi: 10.7554/eLife.66481

Figure Lengend Snippet: ( A ) Schematic of the human bladder-chip with co-culture of the 5637 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 hr. The cycle consisted of a linear increase in strain through stretching of the membrane ( filling bladder , 0–2 hr), maintenance of the membrane under stretch ( filled bladder , 2–4 hr), a quick relaxation of applied strain over 2 min ( voiding bladder , 4:02 hr) and maintenance without applied strain ( voided bladder , 4:02 hr to 6 hr). ( 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: Other , Pooled human female urine , Golden West Diagnostics , Cat#: OH2010-pH , .

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

Journal: eLife

Article Title: Dynamic persistence of UPEC intracellular bacterial communities in a human bladder-chip model of urinary tract infection

doi: 10.7554/eLife.66481

Figure Lengend Snippet:

Article Snippet: Other , Pooled human female urine , Golden West Diagnostics , Cat#: OH2010-pH , .

Techniques: Isolation, Recombinant, Plasmid Preparation, Saline, Modification, Clinical Proteomics, Membrane, Staining, Software