human fibroblasts hff 1 treatment  (ATCC)

Journal: Applied Biological Chemistry

Article Title: Anti-aging potential of Cephalotaxus harringtonia extracts: the role of harringtonine and homoharringtonine in skin protection

doi: 10.1186/s13765-024-00951-1

Figure Lengend Snippet: Fig. 5 Survival of normal human dermal fibroblasts (NHDFs) under treatment with harringtonine (1), homoharringtonine (2) and ginkgetin (3). NHDFs were seeded at 1 × 104 cells/well in 96-well plates, and incubated for 24 h, after which, NHDFs were exposed to each concentration of samples. After 24 h, cell viability was measured using an EZ-Cytox solution. Cell viability results are presented as a percentage of the vehicle control. The experiments were conducted in triplicate and the graphs present the mean ± SEM

Article Snippet: Cell culture and treatment preparation Normal human dermal fibroblasts (NHDFs) (juvenile foreskin-derived, cryopreserved, 1 × 104 cells) were obtained from PromoCell (Sickingenstr, Heidelberg, Germany) and stored in a nitrogen tank.

Techniques: Incubation, Concentration Assay, Control

Journal: Applied Biological Chemistry

Article Title: Anti-aging potential of Cephalotaxus harringtonia extracts: the role of harringtonine and homoharringtonine in skin protection

doi: 10.1186/s13765-024-00951-1

Figure Lengend Snippet: Fig. 6 Effects of extracts CH-10Y-buds, CH-10Y-leaves, CH-200Y-buds, and CH-200Y-leaves on intracellular ROS accumulation in TNF-α stimulated normal human dermal fibroblasts (NHDFs). (A) NHDFs were seeded at 1 × 104 cells/well in 96-well black plates and incubated for 24 h. (B) NHDFs were seeded at 2 × 104 cells/well in 48-well plates and incubated for 24 h. The medium was replaced with a serum-free medium to create starvation conditions. After 24 h, the cells were treated with the samples for 1 h, followed by exposure to TNF-α for 15 min. The cells were stained with DCFDA for 15 min and their fluorescence was measured using a Plate Reader (SPARK 10 M; Tecan) at wavelengths of 485 and 535 nm. and fluorescence microscope (Olympus, Tokyo, Japan). The intracellular ROS levels are presented as a percentage of the vehicle control. The results were obtained from three replicate experiments, and the graphs are presented as mean ± SEM. ###p < 0.001 vs. vehicle control. *p < 0.05, and ***p < 0.001 vs. TNF-α stimulated control

Article Snippet: Cell culture and treatment preparation Normal human dermal fibroblasts (NHDFs) (juvenile foreskin-derived, cryopreserved, 1 × 104 cells) were obtained from PromoCell (Sickingenstr, Heidelberg, Germany) and stored in a nitrogen tank.

Techniques: Incubation, Staining, Fluorescence, Microscopy, Control

Journal: Applied Biological Chemistry

Article Title: Anti-aging potential of Cephalotaxus harringtonia extracts: the role of harringtonine and homoharringtonine in skin protection

doi: 10.1186/s13765-024-00951-1

Figure Lengend Snippet: Fig. 7 Survival of normal human dermal fibroblasts (NHDFs) under treatment with harringtonine (1), homoharringtonine (2) and ginkgetin (3). NHDFs were seeded at 1 × 104 cells/well in 96-well plates, and incubated for 24 h, after which, NHDFs were exposed to each concentration of samples. After 24 h, cell viability was measured using an EZ-Cytox solution. Cell viability results are presented as a percentage of the vehicle control. The experiments were conducted in triplicate and the graphs present the mean ± SEM. *p < 0.05, and ***p < 0.001

Article Snippet: Cell culture and treatment preparation Normal human dermal fibroblasts (NHDFs) (juvenile foreskin-derived, cryopreserved, 1 × 104 cells) were obtained from PromoCell (Sickingenstr, Heidelberg, Germany) and stored in a nitrogen tank.

Techniques: Incubation, Concentration Assay, Control

Journal: Applied Biological Chemistry

Article Title: Anti-aging potential of Cephalotaxus harringtonia extracts: the role of harringtonine and homoharringtonine in skin protection

doi: 10.1186/s13765-024-00951-1

Figure Lengend Snippet: Fig. 8 Effects of harringtonine (1), homoharringtonine (2), and ginkgetin (3) on intracellular ROS accumulation, MMP-1 secretion, and type1 procollagen secretion in TNF-α stimulated normal human dermal fibroblasts (NHDFs). (A) NHDFs were seeded at 1 × 104 cells/well in 96-well black plates, incubated for 24 h, and the medium was replaced with a serum-free medium. After 24 h, the cells were treated with the samples for 1 h, and the cells were exposed to 20 ng/mL TNF-α and DCFDA for 15 min. The intracellular ROS is presented as a percent of the vehicle control. (B, C) NHDFs were seeded at 2 × 104 cells/ well in 48-well plates, incubated for 24 h, and the medium was replaced with a serum-free medium. After 24 h, the cells were treated with the samples for 1 h, and the cells were exposed to 20 ng/mL TNF-α for 24 h. MMP-1 secretion was measured in the cell supernatants using an ELISA kit. MMP-1 and col lagen secretion are presented as a fold increase. The results were obtained through two replicate experiments, and the graphs present the mean ± SEM. ##p < 0.01 and ###p < 0.001 vs. vehicle control. *p < 0.05, **p < 0.01, and ***p < 0.001 vs. TNF-α stimulated control

Article Snippet: Cell culture and treatment preparation Normal human dermal fibroblasts (NHDFs) (juvenile foreskin-derived, cryopreserved, 1 × 104 cells) were obtained from PromoCell (Sickingenstr, Heidelberg, Germany) and stored in a nitrogen tank.

Techniques: Incubation, Control, Enzyme-linked Immunosorbent Assay