human skin keratinocytes (hacat) cell lines  (Elabscience Biotechnology)

Journal: Scientific Reports

Article Title: Exploring the potential of 7,4′-di(diethylamino)flavylium as a novel photosensitizer for topical photodynamic therapy of skin cancer

doi: 10.1038/s41598-024-80860-x

Figure Lengend Snippet: Cytotoxicity effects after 24 h incubation of increasing concentrations of 7,4′diN(Et) 2 dye with HaCat, SCC-25, and A375 cells. Results are expressed as the mean ± standard error of the mean (SEM). Two-way analysis of variance (two-way ANOVA) was used to determine statistically significant differences between the means of the three cell lines using Sidak’s multiple comparisons tests (**** p value < 0.0001, *** p value < 0.001, ** p value < 0.01, * p value < 0.05).

Article Snippet: Human skin keratinocyte cell line (HaCat) was provided by CLS Cell Lines Service GmbH and maintained in DMEM High Glucose Basic (820300a, CLS).

Techniques: Incubation

Journal: Scientific Reports

Article Title: Exploring the potential of 7,4′-di(diethylamino)flavylium as a novel photosensitizer for topical photodynamic therapy of skin cancer

doi: 10.1038/s41598-024-80860-x

Figure Lengend Snippet: ( A ) Photodynamic effect of 7,4′diN(Et) 2 dye on cellular viability of HaCat, SCC-25, and A374 after a 15-minute irradiation period (white light, 22.5 J/cm 2 ). Results are expressed as the mean ± standard error of the mean (SEM). Two-way analysis of variance (two-way ANOVA) was used to determine statistically significant differences between the three cell lines using Sidak’s multiple comparisons tests (**** p value < 0.0001, *** p value < 0.001, ** p value < 0.01, * p value < 0.05). ( B ) Dose response curves for each cell line, indicating the corresponding IC 50 (half maximal inhibitory concentration) values.

Article Snippet: Human skin keratinocyte cell line (HaCat) was provided by CLS Cell Lines Service GmbH and maintained in DMEM High Glucose Basic (820300a, CLS).

Techniques: Irradiation, Concentration Assay

Journal: Scientific Reports

Article Title: Exploring the potential of 7,4′-di(diethylamino)flavylium as a novel photosensitizer for topical photodynamic therapy of skin cancer

doi: 10.1038/s41598-024-80860-x

Figure Lengend Snippet: Photodynamic effect of 7,4′diN(Et) 2 dye at 1.5 µM on cellular viability of HaCat, SCC-25 and A374 after a 15-minute irradiation period (white light, 22.5 J/cm 2 ). Cells were either exposed directly to light in the presence of 7,4′diN(Et) 2 (full colored bars) or washed before irradiation (bars with square pattern). Results are expressed as the mean ± standard error of the mean (SEM). Two-way analysis of variance (two-way ANOVA) was used to determine statistically significant differences between the means of different treatment groups (washed vs. unwashed) within each cell line using Sidak’s multiple comparisons tests (**** p value < 0.0001, *** p value < 0.001, ** p value < 0.01, * p value < 0.05).

Article Snippet: Human skin keratinocyte cell line (HaCat) was provided by CLS Cell Lines Service GmbH and maintained in DMEM High Glucose Basic (820300a, CLS).

Techniques: Irradiation

Journal: Scientific Reports

Article Title: Exploring the potential of 7,4′-di(diethylamino)flavylium as a novel photosensitizer for topical photodynamic therapy of skin cancer

doi: 10.1038/s41598-024-80860-x

Figure Lengend Snippet: Uptake efficiency after 30 min of incubation of 7,4′diN(et) 2 with the different cell lines.

Article Snippet: Human skin keratinocyte cell line (HaCat) was provided by CLS Cell Lines Service GmbH and maintained in DMEM High Glucose Basic (820300a, CLS).

Techniques: Incubation, Concentration Assay

Journal: Scientific Reports

Article Title: Exploring the potential of 7,4′-di(diethylamino)flavylium as a novel photosensitizer for topical photodynamic therapy of skin cancer

doi: 10.1038/s41598-024-80860-x

Figure Lengend Snippet: Confocal laser scanning microscopy images of HaCat, SCC-25 and A374 after incubation with 3 µM of 7,4′diN(Et) 2 dye. Cells stained with DAPI (blue signal), cellular accumulation of photosensitizer (red signal), and merged images (right panel) showing the localization of the photosensitizer. Scale bar of 20 μm.

Article Snippet: Human skin keratinocyte cell line (HaCat) was provided by CLS Cell Lines Service GmbH and maintained in DMEM High Glucose Basic (820300a, CLS).

Techniques: Confocal Laser Scanning Microscopy, Incubation, Staining

Journal: Scientific Reports

Article Title: Exploring the potential of 7,4′-di(diethylamino)flavylium as a novel photosensitizer for topical photodynamic therapy of skin cancer

doi: 10.1038/s41598-024-80860-x

Figure Lengend Snippet: Confocal laser scanning microscopy images of HaCat and A374 CLSM images showing colocalization between lysosomes (marked with LumiTracker ® Lyso Green) and 7,4′diN(Et) 2 after incubation for 30 min. The merged images (right panel) show the lysosomal accumulation of the dye (yellow color). Scale bar of 10 μm.

Article Snippet: Human skin keratinocyte cell line (HaCat) was provided by CLS Cell Lines Service GmbH and maintained in DMEM High Glucose Basic (820300a, CLS).

Techniques: Confocal Laser Scanning Microscopy, Incubation

Journal: Scientific Reports

Article Title: Exploring the potential of 7,4′-di(diethylamino)flavylium as a novel photosensitizer for topical photodynamic therapy of skin cancer

doi: 10.1038/s41598-024-80860-x

Figure Lengend Snippet: Confocal laser scanning microscopy images of HaCat, SCC-25 and A374 after being incubated with 3 µM of 7,4′diN(Et) 2 and irradiated (15 min, 22.5 J/cm 2 ). Intracellular ROS generation indicated by the green fluorescence signal of DCDFA. Within each subset of microscopy images, the right panel represents the merge between the DCFA signal and bright field contrast image. Scale bar of 20 μm.

Article Snippet: Human skin keratinocyte cell line (HaCat) was provided by CLS Cell Lines Service GmbH and maintained in DMEM High Glucose Basic (820300a, CLS).

Techniques: Confocal Laser Scanning Microscopy, Incubation, Irradiation, Fluorescence, Microscopy

Journal: Cell reports

Article Title: Oct4 is a gatekeeper of epithelial identity by regulating cytoskeletal organization in skin keratinocytes.

doi: 10.1016/j.celrep.2024.113859

Figure Lengend Snippet: Figure 3. Oct4 impacts actin cytoskeleton organization and fibronectin deposition in the ECM (A) Representative confocal images of WT keratinocytes and OCT4 knockout clones stained with phalloidin for F-actin visualization at 403 magnification. Scale bars, 100 mm. (B) Representative images of WT keratinocytes and OCT4 knockout clones stained with phalloidin at 633 magnification. Magnified area (white rectangle): detailed visualization of stress fibers. White arrows point to stress fibers. Scale bars, 100 mm and 25 mm for magnified images.

Article Snippet: HaCaT immortalized keratinocyte cell line frommale adult human skin (CLS, 300493) and HeLa human cervical cancer cell line (CLS, 300194) were cultured at 37 C in 5% CO2, in DMEM (Gibco, 41965039) supplemented with 10% FBS (Gibco, 10500064) and 1% penicillin-streptomycin (Gibco, 15070063).

Techniques: Knock-Out, Clone Assay, Staining

Journal: Cell reports

Article Title: Oct4 is a gatekeeper of epithelial identity by regulating cytoskeletal organization in skin keratinocytes.

doi: 10.1016/j.celrep.2024.113859

Figure Lengend Snippet: Figure 7. Phenotypic changes in skin keratinocytes are partially dependent on Oct4 transcriptional function (A) Schematic depicting Oct4 pCEP constructs: pCEP-OCT4-WT (top), pCEP-OCT4-AA resembles/mimics the WT construct (center), and the pCEP-OCT4-EE mutant does not bind to DNA (bottom). (B) Representative western blot images showing Oct4 protein expression in OCT4 knockout clones transfected with the constructs mentioned in (A). (C) RT-qPCR quantification of OCT4 mRNA levels, comparing WT cells with OCT4 knockout clones transfected with the constructs mentioned in (A). n = 3, mean ± SEM, one-way ANOVA. (D) Representative phase-contrast images demonstrating changes in cell size and morphology: OCT4 knockout clones left UT or transfected with the pCEP- OCT4-WT, pCEP-OCT4-AA, or pCEP-OCT4-EE construct compared with WT cells. Scale bar, 100 mm. (E) Quantification of cell size, comparing UT OCT4 knockout cells with clones transfected with the pCEP-OCT4-WT, pCEP-OCT4-AA, or pCEP-OCT4-EE construct. n = 3, mean ± SEM, one-way ANOVA. (F) RT-qPCR quantification of WASP, CDC42, and ROCK1 transcript levels in UT clone 1 compared with clone 1 transfected with the pCEP-OCT4-WT, pCEP- OCT4-AA, or pCEP-OCT4-EE construct. n = 3, mean ± SEM, one-way ANOVA. (G) RT-qPCR quantification of WASP, CDC42, and ROCK1 transcript levels in UT clone 3 compared with clone 3 transfected with the pCEP-OCT4-WT, pCEP- OCT4-AA, or pCEP-OCT4-EE construct. n = 3, mean ± SEM, one-way ANOVA. (H) RT-qPCR quantification of WASP, CDC42, and ROCK1 transcript levels in UT clone 4 compared with clone 4 transfected with the pCEP-OCT4-WT, pCEP- OCT4-AA, or pCEP-OCT4-EE construct. n = 3, mean ± SEM, one-way ANOVA. See also Table S2.

Article Snippet: HaCaT immortalized keratinocyte cell line frommale adult human skin (CLS, 300493) and HeLa human cervical cancer cell line (CLS, 300194) were cultured at 37 C in 5% CO2, in DMEM (Gibco, 41965039) supplemented with 10% FBS (Gibco, 10500064) and 1% penicillin-streptomycin (Gibco, 15070063).

Techniques: Construct, Mutagenesis, Western Blot, Expressing, Knock-Out, Clone Assay, Transfection, Quantitative RT-PCR