Normal human urothelial cells (HUC) were purchased from ScienCell. These cells were maintained in urothelial cell medium supplemented with 1% penicillin/streptomycin and urothelial cell growth supplement and incubated in a 5% CO2 humidity environment at 37°C. hTERT expressing human urothelial cells, hTU1, were obtained as a generous gift from Dr. Louis Liou (described in Kim et al., 2011 (link)). hTU1 cells were further subcloned and a stable clone was selected (hTUC1-38). Cells were maintained in keratinocyte-SFM and incubated in a 5% CO2 humidified environment at 37°C. Cells were maintained as adherent subconfluent monolayers, fed twice weekly and subcultured at least once weekly. Cells were routinely tested for mycoplasma contamination.
Normal human urothelial cells
Manufactured by ScienCell
Normal human urothelial cells (HUC) are primary cells derived from the urothelium, the inner lining of the urinary bladder. These cells are useful for studying the normal structure and function of the urothelium.
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2 protocols using normal human urothelial cells
1
Immortalized Human Urothelial Cell Lines
2
Immortalized Normal and IC/PBS Urothelial Cells
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Normal human urothelial cells (HUC) were obtained from ScienCell Research Laboratories (Carlsbad, CA), cell isolations from 3 separate donors were used. Urothelial cells isolated from normal bladder (5 separate donors) and the bladder of patients with IC/PBS (5 separate donors) were immortalized with HPV type 16E6E7as described previously [15 (link), 16 (link)]. Samples were obtained from IC/PBS patients by biopsy or bladder washing during cystoscopy. Samples were collected according to an Oklahoma University Health Sciences Center Institutional Review Board approved protocol following informed written consent.
Urothelial cells were grown in EpiLife Media (Cascade Biologics, Inc. Portland, OR) with calcium (0.06mM), growth factor supplements provided by the manufacturer and penicillin (20U/ml)/streptomycin (100mg/ml) (Sigma Chemical Company, St. Louis, MO). After reaching confluence, cells were stratified in the same medium with 10% fetal bovine serum (FBS) and additional 1.0 mM calcium (Ca/FBS). Experiments were conducted at 1, 3, and 7 days after Ca/ FBS addition. These cells in culture show expression of adherens junctions, tight junctions and claudins [17 (link)].
Urothelial cells were grown in EpiLife Media (Cascade Biologics, Inc. Portland, OR) with calcium (0.06mM), growth factor supplements provided by the manufacturer and penicillin (20U/ml)/streptomycin (100mg/ml) (Sigma Chemical Company, St. Louis, MO). After reaching confluence, cells were stratified in the same medium with 10% fetal bovine serum (FBS) and additional 1.0 mM calcium (Ca/FBS). Experiments were conducted at 1, 3, and 7 days after Ca/ FBS addition. These cells in culture show expression of adherens junctions, tight junctions and claudins [17 (link)].
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