primary normal human epidermal keratinocytes nhek  (PromoCell)

Journal: Lasers in Medical Science

Article Title: Contrasting biological responses of gingival fibroblasts and keratinocyte to blue and violet light irradiation: implications for photobiomodulation use in the therapeutic management of periodontal disease

doi: 10.1007/s10103-026-04817-4

Figure Lengend Snippet: Influence of blue (457 nm, A-D) and violet (418 nm, E-F) light upon the metabolic activity and cytotoxicity of pHGFs (A, C and E, respectively) and pHGKs (B, D and F, respectively). Cells were irradiated with different fluences of blue and violet light. At 24 h post-treatment, the metabolic activity of the cells was assessed using AlamarBlue™. Metabolic activity was expressed as a percentage change compared to untreated controls; where the untreated controls were normalised to 100%, represented by the dashed line. LDH release, a marker of cytotoxicity, was also assessed 24 h post-treatment and cytotoxicity calculated as the percentage change in LDH levels compared to untreated controls, normalised to 0%. N = 3, mean ± SD (*p < 0.05, **p < 0.01, ****p < 0.0001)

Article Snippet: Primary human gingival fibroblasts (pHGFs) and primary human gingival keratinocytes (pHGKs) were sourced from ATCC/LGC Standards (Manassas, Virginia, USA). pHGF cells (healthy 60-year-old Caucasian female donor) were maintained in Dulbecco’s Modified Eagle Medium (DMEM), 10% foetal calf serum (FCS, ThermoFisher Scientific, Paisley, UK), 100 units/mL penicillin G sodium, 0.1 μg/mL streptomycin sulphate and 0.25 μg/mL amphotericin (ThermoFisher Scientific). pHGKs (healthy 65-year-old Caucasian male donor) and were maintained in Dermal Cell Basal Medium (DCBM, ATCC), supplemented with a Keratinocyte Growth Kit (ATCC).

Techniques: Activity Assay, Irradiation, Marker

Journal: Lasers in Medical Science

Article Title: Contrasting biological responses of gingival fibroblasts and keratinocyte to blue and violet light irradiation: implications for photobiomodulation use in the therapeutic management of periodontal disease

doi: 10.1007/s10103-026-04817-4

Figure Lengend Snippet: Assessment of ROS generation following the blue light (457 nm) treatment of pHGFs ( A ) and pHGKs ( B ). ROS levels were determined immediately following blue light treatment and the oxidation of added H 2 DCFDA resulting in the emittance of a green, fluorescent signal. ROS generation is reported as a value proportional to relative fluorescence units (RFU). N = 3, mean ± SD (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001)

Article Snippet: Primary human gingival fibroblasts (pHGFs) and primary human gingival keratinocytes (pHGKs) were sourced from ATCC/LGC Standards (Manassas, Virginia, USA). pHGF cells (healthy 60-year-old Caucasian female donor) were maintained in Dulbecco’s Modified Eagle Medium (DMEM), 10% foetal calf serum (FCS, ThermoFisher Scientific, Paisley, UK), 100 units/mL penicillin G sodium, 0.1 μg/mL streptomycin sulphate and 0.25 μg/mL amphotericin (ThermoFisher Scientific). pHGKs (healthy 65-year-old Caucasian male donor) and were maintained in Dermal Cell Basal Medium (DCBM, ATCC), supplemented with a Keratinocyte Growth Kit (ATCC).

Techniques: Fluorescence

Journal: Lasers in Medical Science

Article Title: Contrasting biological responses of gingival fibroblasts and keratinocyte to blue and violet light irradiation: implications for photobiomodulation use in the therapeutic management of periodontal disease

doi: 10.1007/s10103-026-04817-4

Figure Lengend Snippet: Heatmaps depicting the fold changes in the gene expression of specific antioxidants (defined in Table ), proposed to be upregulated through ROS generation and measured at 6 h ( A , C ) and 24 h ( B , D ) in pHGFs and pHGKs, respectively. Relative gene expression is displayed as colours ranging from black to yellow, according to the scale, where the highest fold change is shown in yellow. Expression of target genes in untreated samples is represented by 1 on the colour scale (N = 3)

Article Snippet: Primary human gingival fibroblasts (pHGFs) and primary human gingival keratinocytes (pHGKs) were sourced from ATCC/LGC Standards (Manassas, Virginia, USA). pHGF cells (healthy 60-year-old Caucasian female donor) were maintained in Dulbecco’s Modified Eagle Medium (DMEM), 10% foetal calf serum (FCS, ThermoFisher Scientific, Paisley, UK), 100 units/mL penicillin G sodium, 0.1 μg/mL streptomycin sulphate and 0.25 μg/mL amphotericin (ThermoFisher Scientific). pHGKs (healthy 65-year-old Caucasian male donor) and were maintained in Dermal Cell Basal Medium (DCBM, ATCC), supplemented with a Keratinocyte Growth Kit (ATCC).

Techniques: Gene Expression, Expressing

Journal: Frontiers in Immunology

Article Title: A skin isolate of Micrococcus luteus negates the Staphylococcus aureus- induced release of type 2 cytokines from keratinocytes

doi: 10.3389/fimmu.2026.1711723

Figure Lengend Snippet: The ability to negate S. aureus -induced cytokine secretion from keratinocytes is a strain specific effect of M. luteus CFCS. (A, B) NHEK were treated with FSA or co-treated with FSA and skin bacterial CFCS for 24 h before quantifying IL-33 and TSLP in cell culture medium using ELISA. Stimulation of NHEK with FSA caused an increase in IL-33 and TSLP release. (B) Co-treatment with skin isolated M. luteus FAML CFCS negated FSA-induced release of IL-33 and TSLP. (C) Co-treatment with the M. luteus type strain NCTC 2665 had no effect on FSA-induced IL-33 and TSLP release. Data are expressed as mean ± SEM (n≥3). P values determined by one-way ANOVA *P ≤ 0.05 **P ≤ 0.01 ***P ≤ 0.001 ****P ≤ 0.0001 compared with FSA treated NHEK; ns, non-significant.

Article Snippet: Primary Normal Human Epidermal Keratinocytes (NHEK) (PromoCell, Heidelberg, Germany) were isolated from the epidermis of juvenile foreskin from pooled donors and grown in KGM supplemented with KGM SupplementMix (Promocell) at 37 ̊C in a humidified atmosphere of 5% CO 2 .

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Isolation

Journal: Frontiers in Immunology

Article Title: A skin isolate of Micrococcus luteus negates the Staphylococcus aureus- induced release of type 2 cytokines from keratinocytes

doi: 10.3389/fimmu.2026.1711723

Figure Lengend Snippet: The efficacious molecule secreted by M. luteus FAML is a putative protein. M. luteus FAML was cultured for 1, 2, 4, 6 and 24 h before harvesting CFCS. NHEK were co-cultured with FSA and M. luteus FAML CFCS for 24 h before measuring (A) TSLP and (B) IL-33 in cell culture medium using ELISA. (C) M . luteus FAML CFCS collected at 24 h lost activity against FSA-induced IL-33 and TSLP release in NHEK after heat treatment (HT) to 85°C. (D) Proteins within M. luteus FAML CFCS were precipitated using acetone, then reconstituted in cell culture medium before testing for activity using the same model. Activity was retained within the protein precipitate (PP). Data are expressed as mean ± SEM (n≥4). P values determined by one-way ANOVA *P ≤ 0.05 **P ≤ 0.01 ***P ≤ 0.001 ****P ≤ 0.0001 compared with FSA treated NHEK; ns, non-significant.

Article Snippet: Primary Normal Human Epidermal Keratinocytes (NHEK) (PromoCell, Heidelberg, Germany) were isolated from the epidermis of juvenile foreskin from pooled donors and grown in KGM supplemented with KGM SupplementMix (Promocell) at 37 ̊C in a humidified atmosphere of 5% CO 2 .

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Activity Assay

Journal: Frontiers in Immunology

Article Title: A skin isolate of Micrococcus luteus negates the Staphylococcus aureus- induced release of type 2 cytokines from keratinocytes

doi: 10.3389/fimmu.2026.1711723

Figure Lengend Snippet: Recombinant PADP negates FSA-induced IL-33 and TSLP release from NHEK. NHEK were co-cultured with FSA and rPADP for 24 h before measuring IL-33 and TSLP in the cell culture medium using ELISA. The rPADP negated FSA-induced IL-33 and TSLP release in NHEK. Data are expressed as mean ± SEM (n≥3). P values determined by one-way ANOVA *P ≤ 0.05 **P ≤ 0.01 ****P ≤ 0.0001 compared with FSA treated NHEK.

Article Snippet: Primary Normal Human Epidermal Keratinocytes (NHEK) (PromoCell, Heidelberg, Germany) were isolated from the epidermis of juvenile foreskin from pooled donors and grown in KGM supplemented with KGM SupplementMix (Promocell) at 37 ̊C in a humidified atmosphere of 5% CO 2 .

Techniques: Recombinant, Cell Culture, Enzyme-linked Immunosorbent Assay