primary normal human epidermal keratinocytes nhek  (PromoCell)

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