fibroblast cell lines hdfn  (ATCC)

Journal: International Microbiology

Article Title: Myconanotechnology: evaluation of Ag-Cu bimetallic nanoparticles synthesized by Ganoderma aff. australe against pathogens and cancer cells

doi: 10.1007/s10123-026-00793-5

Figure Lengend Snippet: Cytotoxic activity of Ganoderma aff. australe aqueous extract and Ag/Cu nanoparticles against cancer and non-cancerous cell lines evaluated by MTT assay. Cells (4,500 cells/well) were seeded in 96-well plates and allowed to adhere for 24 h before treatment. Concentrations tested were based on the amount of aqueous extract used to synthesize nanoparticles (see Table 1 ). All treatments were incubated with cells for 72 h at 37 °C with 5% CO₂. After incubation, cells were rinsed with PBS and incubated with 10 µl MTT solution (5 mg/ml) for 4 h, followed by addition of 100 µl DMSO. Absorbance was measured at 570 nm. Bar graphs show cell viability (expressed as IC₅₀ in mg/ml equivalent of extract) for five cancer cell lines and two non-cancerous control lines. Cancer cell lines: Caco-2 (colon cancer, ATCC HTB-37), HT-29 (colon cancer, ATCC HTN-38), MCF7 (breast cancer, ATCC HTB-22), A-172 (glioblastoma, ATCC CRL-1620), and U-87 MG (glioblastoma, ATCC HTB-14). Non-cancerous control lines: HDFn (human dermal fibroblasts, ATCC PCS-201-010) and Detroit 551 (normal skin fibroblasts, ATCC CCL-110). All cell lines were cultured in DMEM/F12 medium supplemented with 10% FBS, 1% antibiotic-antimycotic, 1% glutamine, 1% nonessential amino acids, and 1% sodium pyruvate. A Aqueous extract (0.5 g/50 ml) showing moderate cytotoxic activity with IC₅₀ values ranging from 1.61 ± 0.35 mg/ml (Caco-2) to 5.78 ± 1.48 mg/ml (Detroit 551), demonstrating baseline bioactivity of fungal metabolites. B M2-3-3 nanoparticles (2 ml extract + 3 ml AgNO₃ + 3 ml CuSO₄) exhibiting the highest cytotoxic efficacy across all cancer cell lines, with particularly remarkable activity against glioblastoma lines A-172 (IC₅₀: 0.26 ± 0.09 mg/ml) and U-87 MG (IC₅₀: 0.31 ± 0.12 mg/ml), and colorectal cancer lines Caco-2 (IC₅₀: 0.39 ± 0.12 mg/ml) and HT-29 (IC₅₀: 0.58 ± 0.28 mg/ml). Critically, M2-3-3 showed selective cytotoxicity with significantly higher IC₅₀ values in non-cancerous lines HDFn (2.87 ± 0.64 mg/ml) and Detroit 551 (3.45 ± 0.89 mg/ml), indicating preferential toxicity toward cancer cells. C M3-2-2 nanoparticles (3 ml extract + 2 ml AgNO₃ + 2 ml CuSO₄) demonstrating intermediate cytotoxic activity with IC₅₀ values consistently higher than M2-3-3 but lower than M5-3-3 across all cancer cell lines. D M5-3-3 nanoparticles (5 ml extract + 3 ml AgNO₃ + 3 ml CuSO₄) showing the lowest cytotoxic activity among the three nanoformulations, though still superior to the crude extract. Data represent mean ± SD of three independent experiments performed in triplicate. Statistical analysis performed using one-way ANOVA followed by Tukey’s HSD post-hoc test. Asterisks indicate significant differences: * p ≤ 0.050, ** p ≤ 0.010, *** p ≤ 0.001. The hierarchical efficacy pattern (M2-3-3 > M3-2-2 > M5-3-3 > crude extract) demonstrates successful nanotechnological enhancement of anticancer properties and establishes M2-3-3 as the optimal formulation with superior therapeutic selectivity. The exceptional sensitivity of glioblastoma cell lines suggests potential application in treating brain cancers, which are notoriously difficult to treat due to blood-brain barrier penetration challenges. Complete IC₅₀ values and statistical comparisons are provided in Table 4

Article Snippet: Fibroblast cell lines HDFn and Detroit 551 (ATCC, PCS-201-010 and CCL-110, respectively) were used to control non-cancerous cells.

Techniques: Activity Assay, MTT Assay, Incubation, Control, Cell Culture, Formulation

Journal: bioRxiv

Article Title: Tissue resident cells differentiate S. aureus from S. epidermidis via IL-1ß following barrier disruption in healthy human skin

doi: 10.1101/2024.02.19.580932

Figure Lengend Snippet: ( A ) Intracellular CFU counts of S. epidermidis 1457 (filled circles) and S. aureus USA300 (empty circles) 3 h p.i. of tape-stripped corneocytes, human keratinocytes (HEKa) and fibroblasts (HDFn). Bars represent means ± SD, dots show individual data points (corneocytes and fibroblasts n=3; keratinocytes n=5; 3 technical replicates). Dotted line = initial inoculum. Significance was determined by a two-way ANOVA with Tukey’s correction for multiple comparisons. Significance is denoted by **** p ≤ 0.0001. ( B ) IL-8 response displayed as x-fold change over uninfected controls measured by ELISA in cell culture supernatants of S. epidermidis 1457 or S. aureus USA300 infected corneocytes, keratinocytes or fibroblasts. Dot plot shows individual data points (corneocytes and fibroblasts n = 3; keratinocytes n = 5; 3 technical replicates each). Significance was determined by a two-way ANOVA with Tukey’s correction for multiple comparisons. Significance is denoted by *** p ≤ 0.001 or **** p ≤ 0.0001. ( C ) Microscopic detection of intracellular S. aureus GFP in cell monolayers of tape-stripped corneocytes, human keratinocytes (HEKa) or fibroblasts (HDFn); uninfected CTR vs 3h infection. Immunostaining of f-actin (red) and nuclei (blue) in HEKa and HDFn, as well as autofluorescence staining of corneocytes (red). Scale bars = 50µm.

Article Snippet: The human dermal fibroblasts cell line (HDFn) (Gibco, Thermo Fischer Scientific Cat# C0045C) was cultured in Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F-12) (Gibco, Thermo Fischer Scientific Cat# 31330038) supplemented with L-glutamine (Thermo Fischer Scientific Cat# A2916801), 15 mM HEPES and 5% charcoal-stripped fetal bovine serum (FBS) (Gibco, Thermo Fischer Scientific Cat# 12676029) at 37°C with 5% CO2.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Infection, Immunostaining, Staining