human bladder cancer urothelial t24 cells  (ATCC)

Journal: PLOS One

Article Title: Integrated light and electron microscopy workflow for morphological, molecular and ultrastructural analysis of spheroids

doi: 10.1371/journal.pone.0342659

Figure Lengend Snippet: Representative haematoxylin–eosin (HE)-stained sections of SV-HUC-1 and T24 spheroids prepared by cryosectioning ( A-F ) and paraffin embedding (G-L) . A necrotic core (indicated by asterisks, A-B , D-E , G-H , J-K ) is visible in most cross sections, except those obtained from peripheral regions. Scale bars: 500 µm (A, D, G, J), 100 µm (B, E, H, K), 50 µm (C, F, I, L).

Article Snippet: Normal human urothelial SV-HUC-1 cells (CRL-9520) and muscle-invasive human bladder cancer urothelial T24 cells (HTB-4) were purchased from ATCC (Manassas, VA, United States) and cultured in a 1:1 mixture of A-DMEM medium (Gibco, Thermo Fisher Scientific, Waltham, MA, United States) and F12 (Sigma-Aldrich, St. Louis, MO, United States), supplemented with 5% fetal bovine serum (Invitrogen, Carlsbad, CA, United States) and 4 mM GlutaMAX (Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques: Staining

Journal: PLOS One

Article Title: Integrated light and electron microscopy workflow for morphological, molecular and ultrastructural analysis of spheroids

doi: 10.1371/journal.pone.0342659

Figure Lengend Snippet: Representative images of paraffin sections show E-cadherin (green) in the plasma membrane in cells of SV-HUC-1 spheroids ( A-a3 ) and N-cadherin (red) in the plasma membrane of T24 spheroids ( B-b3 ). Some SV-HUC-1 cells are also positive for N-cadherin (arrows, a2 and a3 ). Yellow insets (in A and B) are magnified (a1-a3 and b1-b3) and display individual and merged channels of E-cadherin, N-cadherin, and DAPI-stained nuclei. Scale bars: 100 µm (A, B) , 20 µm (a1-a3, b1-b3).

Article Snippet: Normal human urothelial SV-HUC-1 cells (CRL-9520) and muscle-invasive human bladder cancer urothelial T24 cells (HTB-4) were purchased from ATCC (Manassas, VA, United States) and cultured in a 1:1 mixture of A-DMEM medium (Gibco, Thermo Fisher Scientific, Waltham, MA, United States) and F12 (Sigma-Aldrich, St. Louis, MO, United States), supplemented with 5% fetal bovine serum (Invitrogen, Carlsbad, CA, United States) and 4 mM GlutaMAX (Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques: Clinical Proteomics, Membrane, Staining

Journal: PLOS One

Article Title: Integrated light and electron microscopy workflow for morphological, molecular and ultrastructural analysis of spheroids

doi: 10.1371/journal.pone.0342659

Figure Lengend Snippet: SV-HUC-1 (A) and T24 cells (C) form spheroids with a spherical morphology. (B) SV-HUC-1 cells are tightly attached to each other (arrows), and microvilli are seen on their surface. (D) T24 cells are loosely attached, displaying wider intercellular spaces (arrowheads) and have fewer microvilli. Scale bars: 100 µm (A, C) , 10 µm (B, D) .

Article Snippet: Normal human urothelial SV-HUC-1 cells (CRL-9520) and muscle-invasive human bladder cancer urothelial T24 cells (HTB-4) were purchased from ATCC (Manassas, VA, United States) and cultured in a 1:1 mixture of A-DMEM medium (Gibco, Thermo Fisher Scientific, Waltham, MA, United States) and F12 (Sigma-Aldrich, St. Louis, MO, United States), supplemented with 5% fetal bovine serum (Invitrogen, Carlsbad, CA, United States) and 4 mM GlutaMAX (Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques:

Journal: PLOS One

Article Title: Integrated light and electron microscopy workflow for morphological, molecular and ultrastructural analysis of spheroids

doi: 10.1371/journal.pone.0342659

Figure Lengend Snippet: Outermost cells in SV-HUC-1 spheroids display cuboidal morphology (A) , whereas T24 cells are more elongated (B) . The presence of cell junctions in the outermost cell layer of SV-HUC-1 (C) and T24 spheroid (D) (boxed regions) indicates a tight cellular network of the outermost cell layer. The central necrotic zone (asterisks) is filled with necrotic cells in SV-HUC-1 and T24 spheroids (E, F) , as clearly demonstrated on semithin sections (G, H) , prepared before ultrathin sectioning. Scale bars: 100 µm (G-H) , 10 µm (A-B) , 5 µm (E-F) , 1 µm (C-D) .

Article Snippet: Normal human urothelial SV-HUC-1 cells (CRL-9520) and muscle-invasive human bladder cancer urothelial T24 cells (HTB-4) were purchased from ATCC (Manassas, VA, United States) and cultured in a 1:1 mixture of A-DMEM medium (Gibco, Thermo Fisher Scientific, Waltham, MA, United States) and F12 (Sigma-Aldrich, St. Louis, MO, United States), supplemented with 5% fetal bovine serum (Invitrogen, Carlsbad, CA, United States) and 4 mM GlutaMAX (Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques:

Journal: Cancer Genomics & Proteomics

Article Title: Proteomic Signatures of Cellular Reprogramming in Bladder Cancer: Insights into the Acquisition of Cancer Stem-like States and Phenotypic Plasticity

doi: 10.21873/cgp.20578

Figure Lengend Snippet: Reprogramming of HTB-4 bladder cancer cells using SeV-mediated delivery. (A) An illustration of the reprogramming timeline indicates the establishment of iPSC-like cell culture by day 30. (B) Monitoring of the reprogrammed bladder cancer cell line revealed epithelioid morphology and colony-like formations from day 7 onward. After transfer onto vitronectin under feeder-free conditions, the rep HTB-4 cells continue to form bigger stem-cell-like colonies. Colonies were formed by cells that transformed from a mesenchymal to a more epithelial morphology, similar to induced pluripotent stem cells (indicated by red circles). (C) Validation of pluripotent-like cells from bladder cancer cell line HTB-4 by immunofluorescence. Reprogrammed HTB-4 cells exhibited positive staining for pluripotency nuclear markers NANOG, OCT4 and SOX2 and cell surface markers TRA-1-60, SSEA-4 and TRA-1-81, while parental HTB-4 cells failed to express these markers—cell nuclei stained with DAPI. Scale bars represent 100 μm.

Article Snippet: HTB-4 cells were grown in DMEM F12K (ATCC), supplemented with 10% FBS (Thermo Fisher Scientific, Waltham, MA, USA) 1% Penicillin/Streptomycin (Capricorn Scientific, Frankfurt, Germany), and 1% L-Glutamine (Capricorn Scientific).

Techniques: Cell Culture, Transformation Assay, Biomarker Discovery, Immunofluorescence, Staining

Journal: Cancer Genomics & Proteomics

Article Title: Proteomic Signatures of Cellular Reprogramming in Bladder Cancer: Insights into the Acquisition of Cancer Stem-like States and Phenotypic Plasticity

doi: 10.21873/cgp.20578

Figure Lengend Snippet: Behavioral assays were used to compare the parental and reprogrammed bladder cancer cell migration and clonogenicity. (A) Cell migration in reprogrammed HTB-4 cells was slower compared to the control group. (B) Colony-forming ability was reduced in the reprogrammed HTB-4 cells in comparison to the parental HTB-4 cells. Unpaired t-test, n=3, *p<0.05. Error bars represent standard deviation. (C) Paclitaxel treatment did not result in a dramatic inhibition of cell migration in parental HTB-4 cells. At higher doses, cell migration could not be evaluated due to cell toxicity. Control HTB-4 vs. treated groups, *p<0.001; control rep HTB-4 vs. treated groups, #p<0.001. (D) Paclitaxel treatment suppressed cell migration in a dose-dependent manner in the Rep HTB-4 cells. (E) The colony-forming ability of parental HTB-4 cells was suppressed by paclitaxel treatment in a dose-dependent manner. Cells exhibited extended, migratory morphology within the colonies. Paclitaxel treatment suppressed the colony-forming ability of rep HTB-4 cells. Cells within the colonies exhibited a more cuboidal and epithelioid morphology. Rep HTB-4 colonies exhibit distinct colony forms both at macroscopic and microscopic levels. Error bars represent standard deviation. One-way ANOVA followed by Tukey’s test, n=2; control HTB-4 vs. treated groups, *p<0.001; control rep HTB-4 vs. treated groups, #p<0.001.

Article Snippet: HTB-4 cells were grown in DMEM F12K (ATCC), supplemented with 10% FBS (Thermo Fisher Scientific, Waltham, MA, USA) 1% Penicillin/Streptomycin (Capricorn Scientific, Frankfurt, Germany), and 1% L-Glutamine (Capricorn Scientific).

Techniques: Migration, Control, Comparison, Standard Deviation, Inhibition

Journal: Cancer Genomics & Proteomics

Article Title: Proteomic Signatures of Cellular Reprogramming in Bladder Cancer: Insights into the Acquisition of Cancer Stem-like States and Phenotypic Plasticity

doi: 10.21873/cgp.20578

Figure Lengend Snippet: Heatmap analysis of global and functional protein expression during bladder cancer cell reprogramming. (A) Whole proteome clustering reveals distinct segregation of reprogrammed bladder cancer cell passages and partial convergence toward SV-HUC-1. (B) Proteins associated with cell adhesion exhibit partial restoration of epithelial-like profile. (C) Proteins in cellular developmental process show progressive modulation in early cellular reprogramming. (D) Cell motility-related proteins exhibit reduced expression in later reprogramming passages, suggesting a shift away from a migratory phenotype. (E) Proteins related to epithelial cell differentiation in rep HTB-4 cells demonstrate increased similarity to SV-HUC-1, highlighting the re-establishment of epithelial characteristics. Color scale represents Z-score–normalized abundance values (green=low; red=high).

Article Snippet: HTB-4 cells were grown in DMEM F12K (ATCC), supplemented with 10% FBS (Thermo Fisher Scientific, Waltham, MA, USA) 1% Penicillin/Streptomycin (Capricorn Scientific, Frankfurt, Germany), and 1% L-Glutamine (Capricorn Scientific).

Techniques: Functional Assay, Expressing, Cell Differentiation

Journal: Cancer Genomics & Proteomics

Article Title: Proteomic Signatures of Cellular Reprogramming in Bladder Cancer: Insights into the Acquisition of Cancer Stem-like States and Phenotypic Plasticity

doi: 10.21873/cgp.20578

Figure Lengend Snippet: Comparative proteomic analysis of reprogrammed bladder cancer cells and normal uroepithelial SV-HUC-1 cells. (A) Venn diagrams showing the total numbers of regulated, upregulated, and downregulated proteins shared or uniquely detected at passages p1, p4, and p7. (B) Protein-protein interaction (PPI) network of proteins commonly upregulated in all three passages (p1, p4, p7) in comparison to the normal uroepithelial cell line SVHUC- 1. Nodes represent proteins and edges show experimentally validated interactions. Enriched pathways are indicated in different colors. (C) PPI network of commonly downregulated proteins across rep HTB-4 p1, p4, p7. (D) Gene ontology (GO) analysis showed enrichment of commonly upregulated proteins in rep HTB-4 p1, p4, and p7 cells, involved in biological processes, molecular function, and cellular components [fold change >2 and false discovery rate (FDR)<0.01]. Significant overrepresentation of processes related to biosynthesis, peptide metabolic pathways, RNA binding, and cadherin binding was detected. (E) Gene ontology (GO) analysis showed enrichment of commonly downregulated proteins in rep HTB-4 p1, p4, and p7 cells, involved in biological processes, molecular function, and cellular components [fold change >2 and false discovery rate (FDR) <0.01]. Reductions in pathways linked to RNA processing, the catenin complex, and the ribonucleoprotein complex were detected.

Article Snippet: HTB-4 cells were grown in DMEM F12K (ATCC), supplemented with 10% FBS (Thermo Fisher Scientific, Waltham, MA, USA) 1% Penicillin/Streptomycin (Capricorn Scientific, Frankfurt, Germany), and 1% L-Glutamine (Capricorn Scientific).

Techniques: Comparison, RNA Binding Assay, Binding Assay

Journal: Cancer Genomics & Proteomics

Article Title: Proteomic Signatures of Cellular Reprogramming in Bladder Cancer: Insights into the Acquisition of Cancer Stem-like States and Phenotypic Plasticity

doi: 10.21873/cgp.20578

Figure Lengend Snippet: Differentially regulated proteins at early reprogramming stages of bladder cancer. (A) STRING analysis of rep HTB-4 p1 cells showing PPI networks of differentially expressed proteins in comparison to parental HTB-4 cells. Enriched GO terms in the cellular component, biological process, and molecular function categories highlight pathways related to membrane organization, cytoskeletal regulation, and cell adhesion. (B) PPI network analysis shows proteins differentially regulated in rep HTB-4 p4 cells. GO enrichment analysis identifies protein cluster enrichments in translation, RNA processing, cytoskeletal regulation and metabolic pathways. (C) PPI network of proteins was significantly regulated at p7 or rep HTB-4 cells compared to parental HTB-4 cells. Functional enrichment analysis shows GO categories of cellular component, molecular function and biological process, including pathways related to cellular respiration, TCA cycle and metabolic remodeling characteristic of later stages during reprogramming. Nodes represent proteins and edges indicate known interactions.

Article Snippet: HTB-4 cells were grown in DMEM F12K (ATCC), supplemented with 10% FBS (Thermo Fisher Scientific, Waltham, MA, USA) 1% Penicillin/Streptomycin (Capricorn Scientific, Frankfurt, Germany), and 1% L-Glutamine (Capricorn Scientific).

Techniques: Comparison, Membrane, Functional Assay

Journal: Translational Oncology

Article Title: Improving photodynamic therapy efficacy in bladder cancer using polymer micelle-encapsulated pheophorbide a

doi: 10.1016/j.tranon.2026.102687

Figure Lengend Snippet: SW780 bladder cancer cells produce floating microbladder-like vesicles (luminal cavities characteristic of urothelial differentiation) in monolayer culture. A. Phase-contrast microscopy of T24 and SW780 bladder cancer cells. B. Scanning electron microscopy (SEM) images showing cell surface morphology. Red arrows indicate floating cyst-like vesicles (microbladder-like, luminal cavities characteristic of urothelial differentiation) produced by the SW780 cell line. Images are representative of the overall cell population.

Article Snippet: Human BC cell lines T24 (ATCC® HTB-4TM, derived from a grade 3 transitional cell carcinoma of an 81-year-old woman) and SW780 (ATCC® CRL-2169TM, grade 1 transitional cell carcinoma from an 80-year-old woman) were purchased in 2022 from ATCC and cultured in Dulbecco’s Modified Eagles Medium containing GlutaMAX supplemented with 10 % of heat-inactivated fetal calf serum, 100 U.mL -1 of penicillin, and 100 μg.mL -1 of streptomycin at 37 °C in a humidified atmosphere at 5 % CO 2 (Panasonic, MCO-170AiCUVH).

Techniques: Microscopy, Electron Microscopy, Produced

Journal: Translational Oncology

Article Title: Improving photodynamic therapy efficacy in bladder cancer using polymer micelle-encapsulated pheophorbide a

doi: 10.1016/j.tranon.2026.102687

Figure Lengend Snippet: Encapsulation of pheophorbide a enhances PDT efficacy in 3D tumor spheroids. A. Scanning electron microscopy (SEM) images showing the surface morphology of T24 and SW780 spheroids. B. General appearance of bladder tumor spheroids before treatment. Red arrows indicate detached cyst-like vesicles, i.e. “microbladders” (luminal cavities characteristic of urothelial differentiation); dotted circles highlight internal cyst-like structures embedded within the spheroids. C. Counterstained semi-thin transverse sections (500 nm) of spheroids revealing internal cyst-like structures (dotted circles). D. Two-photon microscopy imaging of pheophorbide a (Pheo) (1 µM) penetration in T24 spheroids after 30 min incubation at 37 °C either in its free or encapsulated formulation. Poly (ethylene oxide)-block-poly (ε-caprolactone) (PEO 5000 -PCL 4000 ) empty micelles; Pheophorbide encapsulated in PEO-PCL micelles (Pheo-PEOPCL). Cyan: nuclei (Hoechst); red: pheophorbide fluorescence. E. Spheroid viability assessed by intracellular ATP quantification at 3- and 6-days post-PDT ([Pheo] = 3 µM). Results include data from 1 to 6 independent experiments (N), with a cumulative number of biological replicates (n) ranging from 6 to 39.

Article Snippet: Human BC cell lines T24 (ATCC® HTB-4TM, derived from a grade 3 transitional cell carcinoma of an 81-year-old woman) and SW780 (ATCC® CRL-2169TM, grade 1 transitional cell carcinoma from an 80-year-old woman) were purchased in 2022 from ATCC and cultured in Dulbecco’s Modified Eagles Medium containing GlutaMAX supplemented with 10 % of heat-inactivated fetal calf serum, 100 U.mL -1 of penicillin, and 100 μg.mL -1 of streptomycin at 37 °C in a humidified atmosphere at 5 % CO 2 (Panasonic, MCO-170AiCUVH).

Techniques: Encapsulation, Electron Microscopy, Microscopy, Imaging, Incubation, Formulation, Blocking Assay, Fluorescence

Journal: Translational Oncology

Article Title: Improving photodynamic therapy efficacy in bladder cancer using polymer micelle-encapsulated pheophorbide a

doi: 10.1016/j.tranon.2026.102687

Figure Lengend Snippet: Encapsulation of pheophorbide a in PEO-PCL micelles enhances PDT-induced cytotoxicity in 2D high-grade (T24) and low-grade (SW780) bladder cancer cell cultures. A. Cells were treated with empty poly (ethylene oxide)-block-poly (ε-caprolactone) PEO-PCL micelles (PEOPCL) at polymer concentrations equivalent to those used in pheo-loaded micelles, based on the 1:30 drug-to-polymer weight ratio. B. Cells were exposed to increasing concentrations of free pheophorbide a (Pheo) (nM). C. Cells were treated with increasing concentrations of Pheo encapsulated in PEO-PCL micelles (pheo–PEOPCL) (nM). D. Half-maximal inhibitory concentration (IC₅₀) determination for the Pheo–PEOPCL condition. Cell confluence was monitored by videomicroscopy for 72 h post-PDT ( N > 1; n = 6).

Article Snippet: Human BC cell lines T24 (ATCC® HTB-4TM, derived from a grade 3 transitional cell carcinoma of an 81-year-old woman) and SW780 (ATCC® CRL-2169TM, grade 1 transitional cell carcinoma from an 80-year-old woman) were purchased in 2022 from ATCC and cultured in Dulbecco’s Modified Eagles Medium containing GlutaMAX supplemented with 10 % of heat-inactivated fetal calf serum, 100 U.mL -1 of penicillin, and 100 μg.mL -1 of streptomycin at 37 °C in a humidified atmosphere at 5 % CO 2 (Panasonic, MCO-170AiCUVH).

Techniques: Encapsulation, Blocking Assay, Polymer, Concentration Assay

Journal: Translational Oncology

Article Title: Improving photodynamic therapy efficacy in bladder cancer using polymer micelle-encapsulated pheophorbide a

doi: 10.1016/j.tranon.2026.102687

Figure Lengend Snippet: Reduced wound closure in bladder cancer cell monolayers following photodynamic treatment with encapsulated pheophorbide a . A. Kinetics of wound closure over 24 h post-PDT in T24 (grade 3) and SW780 (grade 1) monolayers, monitored by videomicroscopy. Cells were treated with increasing concentrations of pheophorbide a encapsulated in poly (ethylene oxide)-block-poly (ε-caprolactone) PEO-PCL micelles (Pheo-PEO-PCL) (nM). B. Quantification of wound closure ( %) at 24 h post-PDT across the different Pheo-PEO-PCL concentrations. Relative wound density ( %) reflects the proportion of the scratch area repopulated by migrating cells. Data are expressed as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s multiple comparisons test versus control (0 nM).

Article Snippet: Human BC cell lines T24 (ATCC® HTB-4TM, derived from a grade 3 transitional cell carcinoma of an 81-year-old woman) and SW780 (ATCC® CRL-2169TM, grade 1 transitional cell carcinoma from an 80-year-old woman) were purchased in 2022 from ATCC and cultured in Dulbecco’s Modified Eagles Medium containing GlutaMAX supplemented with 10 % of heat-inactivated fetal calf serum, 100 U.mL -1 of penicillin, and 100 μg.mL -1 of streptomycin at 37 °C in a humidified atmosphere at 5 % CO 2 (Panasonic, MCO-170AiCUVH).

Techniques: Blocking Assay, Control

Journal: Translational Oncology

Article Title: Improving photodynamic therapy efficacy in bladder cancer using polymer micelle-encapsulated pheophorbide a

doi: 10.1016/j.tranon.2026.102687

Figure Lengend Snippet: Proof of concept of photodynamic therapy efficacy in a complex 3D engineered bladder tumor model. A. Histological cross-sections of the vesical reconstructed tissues stained with Masson’s trichrome, 72 hours after PDT with 3 µM of either free pheophorbide a (Pheo) or pheophorbide a encapsulated in poly (ethylene oxide)-block-poly (ε-caprolactone) PEO-PCL micelles (Pheo–PEO-PCL). Cells appear in red, and stromal collagens in blue. Tumor cells are outlined with black dotted lines. Representative images are shown. B. Tissue viability assessed using the PrestoBlue assay at 48- and 72-hours post-PDT on engineered bladder substitutes implanted with T24 or SW780 spheroids. Results are expressed as a percentage of the untreated control at each timepoint ± SEM.

Article Snippet: Human BC cell lines T24 (ATCC® HTB-4TM, derived from a grade 3 transitional cell carcinoma of an 81-year-old woman) and SW780 (ATCC® CRL-2169TM, grade 1 transitional cell carcinoma from an 80-year-old woman) were purchased in 2022 from ATCC and cultured in Dulbecco’s Modified Eagles Medium containing GlutaMAX supplemented with 10 % of heat-inactivated fetal calf serum, 100 U.mL -1 of penicillin, and 100 μg.mL -1 of streptomycin at 37 °C in a humidified atmosphere at 5 % CO 2 (Panasonic, MCO-170AiCUVH).

Techniques: Staining, Blocking Assay, Prestoblue Assay, Control