human skin fibroblast cell line hs68  (ATCC)

Journal: International journal of molecular sciences

Article Title: The Role of Cyclic Adenosine Monophosphate (cAMP) in Modulating Glucocorticoid Receptor Signaling and Its Implications on Glucocorticoid-Related Collagen Loss.

doi: 10.3390/ijms241210180

Figure Lengend Snippet: Figure 5. cAMP-mediated stress reduction improves collagen levels in human dermal fibroblasts. Human dermal fibroblast Hs68 cells were co-transfected with the GRE plasmid and the pRL-TK vector for normalization control. (A) After 24 h, the cells were treated with 1 µM dexamethasone for 1, 5, 12, and 24 h. The cells were then harvested, and a GRE luciferase assay was conducted. (B) Transfected cells were treated with dexamethasone alone (1 µM), forskolin alone (1 µM), or a combination of dexamethasone and forskolin. After incubation for 5 or 24 h, the cells were lysed, and a GRE luciferase assay was performed. (C) Hs68 cells were exposed to dexamethasone alone (1 µM), forskolin alone (1 µM), or dexamethasone and forskolin together for 24, 48, or 72 h. The culture supernatants of dermal fibroblasts were collected daily to measure the content of procollagen type I C-peptide using ELISA. (D) Additionally, 24 h after treatment, the gene expression levels of collagen type I α1 chain (COL1A1) in the cells were quantitated using qRT-PCR. (E) Hs68 cells exposed to dexamethasone were initially exposed to µM PD98059 (50 µM; indicated in purple) and treated with forskolin (1 µM) for 24 h. Subsequently, the content of procollagen type I C-peptide was measured using ELISA after 24 h. The values are presented as the mean ± SEM of three biological replicates (* p < 0.05; ns, not significant).

Article Snippet: HEK293 cells and the human skin fibroblast (Hs68) cell line were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Transfection, Plasmid Preparation, Control, Luciferase, Incubation, Enzyme-linked Immunosorbent Assay, Gene Expression, Quantitative RT-PCR

Journal: Molecules (Basel, Switzerland)

Article Title: Valorization of the Photo-Protective Potential of the Phytochemically Standardized Olive ( Olea europaea L.) Leaf Extract in UVA-Irradiated Human Skin Fibroblasts.

doi: 10.3390/molecules27165144

Figure Lengend Snippet: Figure 2. The effects of different concentrations of OLE on the viability of human skin fibroblasts. Cells cultured (24 h) in the presence of OLE (2.5–25 µg/mL), vehicle (0), or the reference antioxidants AA and QU (25 µg/mL) not irradiated (A) or exposed to UVA-irradiation (8 J/cm2) (B) were cultured for an additional 24 h, and then subjected to CCK-8 assay. The cytotoxicity of OLE and fibroblast survival were expressed as a percentage of control, where the absorbance of untreated and of corresponding non-irradiated cells were taken as 100%, respectively. The post-radiation viability of fibroblasts evaluated by the Annexin V-FITC/PI staining (flow cytometry) assay (C). A representative dot-plot histogram is shown (D), cell populations in II, I and III/IV quadrants represent alive, undergoing necrosis and undergoing apoptosis Hs68 cells, respectively. The figure shows mean results from 3–6 independent experiments. Error bars denote ±SD. * p < 0.05; ** p < 0.01; *** p < 0.001 compared to control (0).

Article Snippet: Human skin (foreskin) fibroblast cell line (Hs68; ATCC® CRL-1635TM) was purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Cell Culture, Irradiation, CCK-8 Assay, Control, Staining, Flow Cytometry