human pancreatic carcinoma cell lines panc1  (ATCC)

Journal: Stem Cell Reviews and Reports

Article Title: Nestin and SOX2 Maintain self-renewal Abilities of Different Pancreatic Cancer Stem Cell Populations

doi: 10.1007/s12015-025-11006-3

Figure Lengend Snippet: Confirmation of siRNA-mediated KD of NES expression in Panc1 cell variants and SOX2 expression in Panc89 cell variants. 5 × 10 4 cells were subjected to siRNA-mediated KD of NES in Panc1 cell variants, SOX2 in Panc89 cell variants and double KD of NES/SOX2 in all cell variants or to CTRLsi transfection for 72 h. Afterward, the KD was confirmed by RT-qPCR (a, c, e, g) and IFS (b, d, f, h). Every analysis was performed with n = 3 independent experiments. Gene expression data of NES and SOX2 (a, c, e, g) are normalized to CTRLsi conditions. All data sets were tested for normal distribution using Shapiro-Wilk test, followed by one-way ANOVA for statistical significance. Significances are indicated by asterisk: p ≤ 0.033 = *, p ≤ 0.002 = **, p ≤ 0.001 = ***. Data are presented by mean with SD. Representative IFS images from n = 3 independent experiments are shown (b, d, f, h). Scale bars IFS = 1000 μm. (KD = knockdown; RT-qPCR = real time quantitative polymerase chain reaction, IFS = immunofluorescence staining, CTRLsi = control siRNA; SD = standard deviation; Holo = Holoclone)

Article Snippet: As a model for mesenchymal-like PDAC cells, the human cell line Panc1 was used (ATCC, Manassas, Virginia, US) originating from a primary tumor of a male PDAC patient.

Techniques: Expressing, Transfection, Quantitative RT-PCR, Gene Expression, Knockdown, Real-time Polymerase Chain Reaction, Immunofluorescence, Staining, Control, Standard Deviation

Journal: Stem Cell Reviews and Reports

Article Title: Nestin and SOX2 Maintain self-renewal Abilities of Different Pancreatic Cancer Stem Cell Populations

doi: 10.1007/s12015-025-11006-3

Figure Lengend Snippet: Gene expression analysis of EMT markers and plasticity modulators after single and double KD of NES and SOX2 in Panc1 and Panc89 cell variants. 5 × 10 4 cells were subjected to siRNA-mediated KD of NES in Panc1 cell variants (a), SOX2 in Panc89 cell variants (c), double KD of NES/SOX2 in Panc1 cell variants (b) and Panc89 cell variants (d) or to CTRLsi transfection (a-d) for 72 h. Afterward, gene expression of EMT markers (a-d top: CDH1 , L1CAM , VIM ) and plasticity modulators (a-d bottom: ZEB1 , ZEB2 , OVOL2 ) was determined via RT-qPCR. Analysis was performed with n = 3 independent experiments. Gene expression data of all genes of interest were first normalized to the reference gene GAPDH and then normalized to CTRLsi conditions. All data sets were tested for normal distribution using Shapiro-Wilk test. Parametric data were tested by one-way ANOVA and are presented by mean with SD. Non-parametric data were tested by Kruskal-Wallis one-way ANOVA on ranks and are presented as median with interquartile range. Significances are indicated by asterisk: p ≤ 0.033 = *, p ≤ 0.002 = **, p ≤ 0.001 = ***. (CSC = cancer stem cell; EMT = Epithelial-to-Mesenchymal Transition; KD = knockdown; RT-qPCR = real time quantitative polymerase chain reaction; CTRLsi = control siRNA; SD = standard deviation; Holo = Holoclone)

Article Snippet: As a model for mesenchymal-like PDAC cells, the human cell line Panc1 was used (ATCC, Manassas, Virginia, US) originating from a primary tumor of a male PDAC patient.

Techniques: Gene Expression, Transfection, Quantitative RT-PCR, Knockdown, Real-time Polymerase Chain Reaction, Control, Standard Deviation

Journal: Stem Cell Reviews and Reports

Article Title: Nestin and SOX2 Maintain self-renewal Abilities of Different Pancreatic Cancer Stem Cell Populations

doi: 10.1007/s12015-025-11006-3

Figure Lengend Snippet: KD of NES in Panc1 cell variants and KD of SOX2 in Panc89 cell variants marginally impact cell growth but significantly decrease self-renewing properties. 5 × 10 4 cells were either left untreated (a) or were subjected to siRNA-mediated KD of NES in Panc1 cell variants (b, d), SOX2 in Panc89 cell variants (c, e), double KD of NES/SOX2 in all cell variants (c-e) or to CTRLsi transfection (b-e) for 72 h. Afterward, the number of viable cells was determined by Trypan blue staining (a-c) and cells were re-seeded with 400 cells/well for Panc1 cell variants or 200 cells/well for Panc89 cell variants for CFA in 12-well plates (d-e). The number of viable cells under KD of NES (b), SOX2 (b) or NES / SOX2 (c) was normalized to CTRLsi conditions. Data were tested for normal distribution by Shapiro-Wilk analysis, followed by one-way ANOVA and Tukey´s multiple comparison test. d-e) CFAs were monitored for 6–10 d, fixed with PFA and stained with crystal violet. CFA data were analyzed by two-way ANOVA and Tukey´s multiple comparison. Every analysis was performed with n = 3 independent experiments plus technical replicates and data are shown as mean with SEM. Significances are indicated by asterisk: p ≤ 0.033 = *, p ≤ 0.002 = **, p ≤ 0.001 = ***. (KD = knockdown; CTRLsi = control siRNA; CFA = colony formation assay; SEM = standard error of means; Holo = Holoclone; PFA = paraformaldehyde; SEM = standard error of means; Holo = Holoclone; No. = Number; w/o KD = without knockdown)

Article Snippet: As a model for mesenchymal-like PDAC cells, the human cell line Panc1 was used (ATCC, Manassas, Virginia, US) originating from a primary tumor of a male PDAC patient.

Techniques: Transfection, Staining, Comparison, Knockdown, Control, Colony Assay

Journal: Stem Cell Reviews and Reports

Article Title: Nestin and SOX2 Maintain self-renewal Abilities of Different Pancreatic Cancer Stem Cell Populations

doi: 10.1007/s12015-025-11006-3

Figure Lengend Snippet: siRNA-mediated KD of NES in Panc1 and SOX2 in Panc89 cell variants marginally impacts migration and invasion properties. 5 × 10 4 cells were subjected to siRNA-mediated KD of NES in Panc1 and SOX2 Panc89 cell variants or to CTRLsi transfection for 72 h. Afterward, transfected cells were detached and seeded in (a) Ibidi ® 2-well culture inserts (4 × 10 4 for Panc1; 3 × 10 4 for Panc89 cell variants) and migration was analyzed after 24 h for Panc1 and Panc89 cell variants, or in (b) 96-well ULA plates for spheroid formation (1 × 10 4 for Panc1 cell variants; 1.5 × 10 4 for Panc89 cell variants) and determination of invasion properties. Cell invasion (number of invasive fronts [#]; invasion distance [µm]) was analyzed after 72 and 120 h for Panc1 and Panc89 cell variants, respectively. Every analysis was performed with n = 3 independent experiments plus technical replicates and NES and SOX2 KD values, respectively, were normalized to CTRLsi conditions. All data sets were tested for normal distribution using Shapiro-Wilk test. Parametric data were tested by one-way ANOVA with Tukey´s multiple comparison and are presented by mean with SEM. Non-parametric data were tested by Kruskal-Wallis one-way ANOVA on ranks and are presented as median with interquartile range. Representative images from n = 3 independent experiments are shown (a, b; top). Scale bar a) = 200 μm, b) left = 400 μm, b) right = 200 μm. (KD = knockdown; CTRLsi = control siRNA; ULA = ultra-low-attachment plate; SEM = standard error of means; Holo = Holoclone)

Article Snippet: As a model for mesenchymal-like PDAC cells, the human cell line Panc1 was used (ATCC, Manassas, Virginia, US) originating from a primary tumor of a male PDAC patient.

Techniques: Migration, Transfection, Comparison, Knockdown, Control

Journal: Stem Cell Reviews and Reports

Article Title: Nestin and SOX2 Maintain self-renewal Abilities of Different Pancreatic Cancer Stem Cell Populations

doi: 10.1007/s12015-025-11006-3

Figure Lengend Snippet: siRNA-mediated KD of NES in Panc1 cell variants and SOX2 in Panc89 cell variants marginally impacts the response to cytostatic drugs. 5 × 10 4 cells were subjected to siRNA-mediated KD of NES in Panc1 and SOX2 Panc89 cell variants or to CTRLsi transfection for 72 h. Afterward, 1 × 10 3 cells/well of either cell variants and conditions were re-seeded in a 96-well plate and after 24 h, the cells were treated with Gemcitabine, 5-FU or Oxaliplatin for 72 h. Then, (a) Panc1 and (b) Panc89 cell variants were stained with Hoechst 33342 (1:5000) and PI (1:50) and imaged to determine the total number of cells (Hoechst-positive, nuclei count) and the number of dead cells (Hoechst/PI-positive, cell death), respectively. Every analysis was performed with n = 3 independent experiments. The nuclei count and cell death values after KD of NES and SOX2 , respectively, were normalized to CTRLsi conditions. All data sets were analyzed by two-way ANOVA and Tukey´s multiple comparison. Every analysis was performed with n = 3 independent experiments plus technical replicates and data are shown as mean with SEM. Significances are indicated by asterisk: p ≤ 0.033 = *, p ≤ 0.002 = **, p ≤ 0.001 = ***. (KD = knockdown; CTRLsi = control siRNA; 5-FU = 5-Fluorouracil; PI = propidium iodide; SEM = standard error of means; Holo = Holoclone)

Article Snippet: As a model for mesenchymal-like PDAC cells, the human cell line Panc1 was used (ATCC, Manassas, Virginia, US) originating from a primary tumor of a male PDAC patient.

Techniques: Transfection, Staining, Comparison, Knockdown, Control