Review



m1 macrophages  (PromoCell)


Bioz Verified Symbol PromoCell is a verified supplier
Bioz Manufacturer Symbol PromoCell manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 95
      Buy from Supplier

    Structured Review

    PromoCell m1 macrophages
    CoMiX-Fc enhances significantly phagocytosis of P. aeruginosa by PBMCs-derived <t>M1</t> <t>macrophages</t> and more importantly by Neutrophils-Like Cells, nevertheless, both CoMiX slightly improve NLCs-dependent antimicrobial activity against the bacteria. PMA-activated M1 macrophages, from four different healthy donors, were co-incubated with pHrodo-stained P. aeruginosa (PAO1) at a 12:1 bacteria-to-cell ratio with 10% NHS and in the presence or absence of 15 μg/mL CoMiX-Fc, CoMiX-FHR1 or CoMiX-irrelevant. Engulfment of bacteria was assessed after 30 min (left panels) and 1 h (right panels) by real-time Incucyte® microscope (a–c) . The percentage of pHrodo positive cells (a) and the intensity of pHrodo rationalised over the surface of cells and calculated as integrated intensity (b) were recorded. Data are presented as the mean values ± SEM. Results correspond to two independent experiments with macrophages from 4 healthy donors (three technical replicates per donor). Statistical analysis was performed using paired one-way ANOVA (to smooth inter-donor variability), followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01. (c) Representative incucyte images for the phagocytosis induced by serum and CoMiX-Fc over time. (d) Fluorescence microscopy image obtained on a wide field Axio Observer Z1, and treated on ImageJ of phagocytosed P. aeruginosa bacteria by PBMCs-derived M1 macrophages in presence of 10% serum and 15 μg/mL CoMiX-Fc. Red = wheat germ agglutinin Alexa-647, staining carbohydates residues of macrophages membranes; green = CellTrace™ CFSE stained P. aeruginosa bacteria. Scale bar = 25 μm. P. aeruginosa PAO1 strain and PAO1-GFP strain were co-cultured with NLCs at a 10:1 bacteria-to-cell ratio, treated with 2% NHS and CoMiX (15 μg/mL) and incubated for 20 min at 37 °C under agitation (200 rpm). After washes and treatment with gentamicin to eliminate non-phagocytosed bacteria from the cells samples were (1) fixed and read by flow cytometry. (e) Gating of NLCs with FSC and SSC to eliminate cell debris and free bacteria from the analysis (left graph). Representative histogram describing the total population of NLCs, and composed of GFP negative cells and GFP positive cells (right graph). To assess phagocytosis, the percentage of GFP positive cells (f) and the mean fluorescence (g) were acquired. Data are presented as the mean values ± SEM. Results correspond to three pooled experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. (h) After washes and treatment with gentamicin, cells were also (2) lysed by Triton X-100, diluted in PBS and plated onto petri dishes. CFUs, corresponding to phagocytosed bacteria were counted in duplicates. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. To assess NLCs-dependent killing, P. aeruginosa PAO1 strain at a MOI of 2:1 was co-cultured with NLCs, treated with 10% NHS and CoMiX (15 μg/mL) for 1 h, plated on petri dishes to count the final bacterial CFUs in duplicates (i) . Conditions which were not co-cultured with NLCs were used as control (100% cell survival). Results are expressed as surviving bacteria compared to bacterial growth under the same conditions in the absence of NLCs. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01.
    M1 Macrophages, supplied by PromoCell, used in various techniques. Bioz Stars score: 95/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m1 macrophages/product/PromoCell
    Average 95 stars, based on 44 article reviews
    m1 macrophages - by Bioz Stars, 2025-12
    95/100 stars

    Images

    1) Product Images from "Directed-complement killing of Pseudomonas aeruginosa protects against lethal pneumonia"

    Article Title: Directed-complement killing of Pseudomonas aeruginosa protects against lethal pneumonia

    Journal: eBioMedicine

    doi: 10.1016/j.ebiom.2025.105926

    CoMiX-Fc enhances significantly phagocytosis of P. aeruginosa by PBMCs-derived M1 macrophages and more importantly by Neutrophils-Like Cells, nevertheless, both CoMiX slightly improve NLCs-dependent antimicrobial activity against the bacteria. PMA-activated M1 macrophages, from four different healthy donors, were co-incubated with pHrodo-stained P. aeruginosa (PAO1) at a 12:1 bacteria-to-cell ratio with 10% NHS and in the presence or absence of 15 μg/mL CoMiX-Fc, CoMiX-FHR1 or CoMiX-irrelevant. Engulfment of bacteria was assessed after 30 min (left panels) and 1 h (right panels) by real-time Incucyte® microscope (a–c) . The percentage of pHrodo positive cells (a) and the intensity of pHrodo rationalised over the surface of cells and calculated as integrated intensity (b) were recorded. Data are presented as the mean values ± SEM. Results correspond to two independent experiments with macrophages from 4 healthy donors (three technical replicates per donor). Statistical analysis was performed using paired one-way ANOVA (to smooth inter-donor variability), followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01. (c) Representative incucyte images for the phagocytosis induced by serum and CoMiX-Fc over time. (d) Fluorescence microscopy image obtained on a wide field Axio Observer Z1, and treated on ImageJ of phagocytosed P. aeruginosa bacteria by PBMCs-derived M1 macrophages in presence of 10% serum and 15 μg/mL CoMiX-Fc. Red = wheat germ agglutinin Alexa-647, staining carbohydates residues of macrophages membranes; green = CellTrace™ CFSE stained P. aeruginosa bacteria. Scale bar = 25 μm. P. aeruginosa PAO1 strain and PAO1-GFP strain were co-cultured with NLCs at a 10:1 bacteria-to-cell ratio, treated with 2% NHS and CoMiX (15 μg/mL) and incubated for 20 min at 37 °C under agitation (200 rpm). After washes and treatment with gentamicin to eliminate non-phagocytosed bacteria from the cells samples were (1) fixed and read by flow cytometry. (e) Gating of NLCs with FSC and SSC to eliminate cell debris and free bacteria from the analysis (left graph). Representative histogram describing the total population of NLCs, and composed of GFP negative cells and GFP positive cells (right graph). To assess phagocytosis, the percentage of GFP positive cells (f) and the mean fluorescence (g) were acquired. Data are presented as the mean values ± SEM. Results correspond to three pooled experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. (h) After washes and treatment with gentamicin, cells were also (2) lysed by Triton X-100, diluted in PBS and plated onto petri dishes. CFUs, corresponding to phagocytosed bacteria were counted in duplicates. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. To assess NLCs-dependent killing, P. aeruginosa PAO1 strain at a MOI of 2:1 was co-cultured with NLCs, treated with 10% NHS and CoMiX (15 μg/mL) for 1 h, plated on petri dishes to count the final bacterial CFUs in duplicates (i) . Conditions which were not co-cultured with NLCs were used as control (100% cell survival). Results are expressed as surviving bacteria compared to bacterial growth under the same conditions in the absence of NLCs. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01.
    Figure Legend Snippet: CoMiX-Fc enhances significantly phagocytosis of P. aeruginosa by PBMCs-derived M1 macrophages and more importantly by Neutrophils-Like Cells, nevertheless, both CoMiX slightly improve NLCs-dependent antimicrobial activity against the bacteria. PMA-activated M1 macrophages, from four different healthy donors, were co-incubated with pHrodo-stained P. aeruginosa (PAO1) at a 12:1 bacteria-to-cell ratio with 10% NHS and in the presence or absence of 15 μg/mL CoMiX-Fc, CoMiX-FHR1 or CoMiX-irrelevant. Engulfment of bacteria was assessed after 30 min (left panels) and 1 h (right panels) by real-time Incucyte® microscope (a–c) . The percentage of pHrodo positive cells (a) and the intensity of pHrodo rationalised over the surface of cells and calculated as integrated intensity (b) were recorded. Data are presented as the mean values ± SEM. Results correspond to two independent experiments with macrophages from 4 healthy donors (three technical replicates per donor). Statistical analysis was performed using paired one-way ANOVA (to smooth inter-donor variability), followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01. (c) Representative incucyte images for the phagocytosis induced by serum and CoMiX-Fc over time. (d) Fluorescence microscopy image obtained on a wide field Axio Observer Z1, and treated on ImageJ of phagocytosed P. aeruginosa bacteria by PBMCs-derived M1 macrophages in presence of 10% serum and 15 μg/mL CoMiX-Fc. Red = wheat germ agglutinin Alexa-647, staining carbohydates residues of macrophages membranes; green = CellTrace™ CFSE stained P. aeruginosa bacteria. Scale bar = 25 μm. P. aeruginosa PAO1 strain and PAO1-GFP strain were co-cultured with NLCs at a 10:1 bacteria-to-cell ratio, treated with 2% NHS and CoMiX (15 μg/mL) and incubated for 20 min at 37 °C under agitation (200 rpm). After washes and treatment with gentamicin to eliminate non-phagocytosed bacteria from the cells samples were (1) fixed and read by flow cytometry. (e) Gating of NLCs with FSC and SSC to eliminate cell debris and free bacteria from the analysis (left graph). Representative histogram describing the total population of NLCs, and composed of GFP negative cells and GFP positive cells (right graph). To assess phagocytosis, the percentage of GFP positive cells (f) and the mean fluorescence (g) were acquired. Data are presented as the mean values ± SEM. Results correspond to three pooled experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. (h) After washes and treatment with gentamicin, cells were also (2) lysed by Triton X-100, diluted in PBS and plated onto petri dishes. CFUs, corresponding to phagocytosed bacteria were counted in duplicates. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. To assess NLCs-dependent killing, P. aeruginosa PAO1 strain at a MOI of 2:1 was co-cultured with NLCs, treated with 10% NHS and CoMiX (15 μg/mL) for 1 h, plated on petri dishes to count the final bacterial CFUs in duplicates (i) . Conditions which were not co-cultured with NLCs were used as control (100% cell survival). Results are expressed as surviving bacteria compared to bacterial growth under the same conditions in the absence of NLCs. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01.

    Techniques Used: Derivative Assay, Activity Assay, Bacteria, Incubation, Staining, Microscopy, Fluorescence, Cell Culture, Flow Cytometry, Control



    Similar Products

    m1 phenotype macrophages  (ATCC)
    98
    ATCC m1 phenotype macrophages
    M1 Phenotype Macrophages, supplied by ATCC, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m1 phenotype macrophages/product/ATCC
    Average 98 stars, based on 1 article reviews
    m1 phenotype macrophages - by Bioz Stars, 2025-12
    98/100 stars
    		  Buy from Supplier
    m1 macrophages  (PromoCell)
    95
    PromoCell m1 macrophages
    CoMiX-Fc enhances significantly phagocytosis of P. aeruginosa by PBMCs-derived <t>M1</t> <t>macrophages</t> and more importantly by Neutrophils-Like Cells, nevertheless, both CoMiX slightly improve NLCs-dependent antimicrobial activity against the bacteria. PMA-activated M1 macrophages, from four different healthy donors, were co-incubated with pHrodo-stained P. aeruginosa (PAO1) at a 12:1 bacteria-to-cell ratio with 10% NHS and in the presence or absence of 15 μg/mL CoMiX-Fc, CoMiX-FHR1 or CoMiX-irrelevant. Engulfment of bacteria was assessed after 30 min (left panels) and 1 h (right panels) by real-time Incucyte® microscope (a–c) . The percentage of pHrodo positive cells (a) and the intensity of pHrodo rationalised over the surface of cells and calculated as integrated intensity (b) were recorded. Data are presented as the mean values ± SEM. Results correspond to two independent experiments with macrophages from 4 healthy donors (three technical replicates per donor). Statistical analysis was performed using paired one-way ANOVA (to smooth inter-donor variability), followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01. (c) Representative incucyte images for the phagocytosis induced by serum and CoMiX-Fc over time. (d) Fluorescence microscopy image obtained on a wide field Axio Observer Z1, and treated on ImageJ of phagocytosed P. aeruginosa bacteria by PBMCs-derived M1 macrophages in presence of 10% serum and 15 μg/mL CoMiX-Fc. Red = wheat germ agglutinin Alexa-647, staining carbohydates residues of macrophages membranes; green = CellTrace™ CFSE stained P. aeruginosa bacteria. Scale bar = 25 μm. P. aeruginosa PAO1 strain and PAO1-GFP strain were co-cultured with NLCs at a 10:1 bacteria-to-cell ratio, treated with 2% NHS and CoMiX (15 μg/mL) and incubated for 20 min at 37 °C under agitation (200 rpm). After washes and treatment with gentamicin to eliminate non-phagocytosed bacteria from the cells samples were (1) fixed and read by flow cytometry. (e) Gating of NLCs with FSC and SSC to eliminate cell debris and free bacteria from the analysis (left graph). Representative histogram describing the total population of NLCs, and composed of GFP negative cells and GFP positive cells (right graph). To assess phagocytosis, the percentage of GFP positive cells (f) and the mean fluorescence (g) were acquired. Data are presented as the mean values ± SEM. Results correspond to three pooled experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. (h) After washes and treatment with gentamicin, cells were also (2) lysed by Triton X-100, diluted in PBS and plated onto petri dishes. CFUs, corresponding to phagocytosed bacteria were counted in duplicates. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. To assess NLCs-dependent killing, P. aeruginosa PAO1 strain at a MOI of 2:1 was co-cultured with NLCs, treated with 10% NHS and CoMiX (15 μg/mL) for 1 h, plated on petri dishes to count the final bacterial CFUs in duplicates (i) . Conditions which were not co-cultured with NLCs were used as control (100% cell survival). Results are expressed as surviving bacteria compared to bacterial growth under the same conditions in the absence of NLCs. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01.
    M1 Macrophages, supplied by PromoCell, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m1 macrophages/product/PromoCell
    Average 95 stars, based on 1 article reviews
    m1 macrophages - by Bioz Stars, 2025-12
    95/100 stars
    		  Buy from Supplier
    m1 macrophage generation medium  (PromoCell)
    95
    PromoCell m1 macrophage generation medium
    CoMiX-Fc enhances significantly phagocytosis of P. aeruginosa by PBMCs-derived <t>M1</t> <t>macrophages</t> and more importantly by Neutrophils-Like Cells, nevertheless, both CoMiX slightly improve NLCs-dependent antimicrobial activity against the bacteria. PMA-activated M1 macrophages, from four different healthy donors, were co-incubated with pHrodo-stained P. aeruginosa (PAO1) at a 12:1 bacteria-to-cell ratio with 10% NHS and in the presence or absence of 15 μg/mL CoMiX-Fc, CoMiX-FHR1 or CoMiX-irrelevant. Engulfment of bacteria was assessed after 30 min (left panels) and 1 h (right panels) by real-time Incucyte® microscope (a–c) . The percentage of pHrodo positive cells (a) and the intensity of pHrodo rationalised over the surface of cells and calculated as integrated intensity (b) were recorded. Data are presented as the mean values ± SEM. Results correspond to two independent experiments with macrophages from 4 healthy donors (three technical replicates per donor). Statistical analysis was performed using paired one-way ANOVA (to smooth inter-donor variability), followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01. (c) Representative incucyte images for the phagocytosis induced by serum and CoMiX-Fc over time. (d) Fluorescence microscopy image obtained on a wide field Axio Observer Z1, and treated on ImageJ of phagocytosed P. aeruginosa bacteria by PBMCs-derived M1 macrophages in presence of 10% serum and 15 μg/mL CoMiX-Fc. Red = wheat germ agglutinin Alexa-647, staining carbohydates residues of macrophages membranes; green = CellTrace™ CFSE stained P. aeruginosa bacteria. Scale bar = 25 μm. P. aeruginosa PAO1 strain and PAO1-GFP strain were co-cultured with NLCs at a 10:1 bacteria-to-cell ratio, treated with 2% NHS and CoMiX (15 μg/mL) and incubated for 20 min at 37 °C under agitation (200 rpm). After washes and treatment with gentamicin to eliminate non-phagocytosed bacteria from the cells samples were (1) fixed and read by flow cytometry. (e) Gating of NLCs with FSC and SSC to eliminate cell debris and free bacteria from the analysis (left graph). Representative histogram describing the total population of NLCs, and composed of GFP negative cells and GFP positive cells (right graph). To assess phagocytosis, the percentage of GFP positive cells (f) and the mean fluorescence (g) were acquired. Data are presented as the mean values ± SEM. Results correspond to three pooled experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. (h) After washes and treatment with gentamicin, cells were also (2) lysed by Triton X-100, diluted in PBS and plated onto petri dishes. CFUs, corresponding to phagocytosed bacteria were counted in duplicates. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. To assess NLCs-dependent killing, P. aeruginosa PAO1 strain at a MOI of 2:1 was co-cultured with NLCs, treated with 10% NHS and CoMiX (15 μg/mL) for 1 h, plated on petri dishes to count the final bacterial CFUs in duplicates (i) . Conditions which were not co-cultured with NLCs were used as control (100% cell survival). Results are expressed as surviving bacteria compared to bacterial growth under the same conditions in the absence of NLCs. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01.
    M1 Macrophage Generation Medium, supplied by PromoCell, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m1 macrophage generation medium/product/PromoCell
    Average 95 stars, based on 1 article reviews
    m1 macrophage generation medium - by Bioz Stars, 2025-12
    95/100 stars
    		  Buy from Supplier
    m1 macrophages inos  (Wanleibio)
    90
    Wanleibio m1 macrophages inos
    CoMiX-Fc enhances significantly phagocytosis of P. aeruginosa by PBMCs-derived <t>M1</t> <t>macrophages</t> and more importantly by Neutrophils-Like Cells, nevertheless, both CoMiX slightly improve NLCs-dependent antimicrobial activity against the bacteria. PMA-activated M1 macrophages, from four different healthy donors, were co-incubated with pHrodo-stained P. aeruginosa (PAO1) at a 12:1 bacteria-to-cell ratio with 10% NHS and in the presence or absence of 15 μg/mL CoMiX-Fc, CoMiX-FHR1 or CoMiX-irrelevant. Engulfment of bacteria was assessed after 30 min (left panels) and 1 h (right panels) by real-time Incucyte® microscope (a–c) . The percentage of pHrodo positive cells (a) and the intensity of pHrodo rationalised over the surface of cells and calculated as integrated intensity (b) were recorded. Data are presented as the mean values ± SEM. Results correspond to two independent experiments with macrophages from 4 healthy donors (three technical replicates per donor). Statistical analysis was performed using paired one-way ANOVA (to smooth inter-donor variability), followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01. (c) Representative incucyte images for the phagocytosis induced by serum and CoMiX-Fc over time. (d) Fluorescence microscopy image obtained on a wide field Axio Observer Z1, and treated on ImageJ of phagocytosed P. aeruginosa bacteria by PBMCs-derived M1 macrophages in presence of 10% serum and 15 μg/mL CoMiX-Fc. Red = wheat germ agglutinin Alexa-647, staining carbohydates residues of macrophages membranes; green = CellTrace™ CFSE stained P. aeruginosa bacteria. Scale bar = 25 μm. P. aeruginosa PAO1 strain and PAO1-GFP strain were co-cultured with NLCs at a 10:1 bacteria-to-cell ratio, treated with 2% NHS and CoMiX (15 μg/mL) and incubated for 20 min at 37 °C under agitation (200 rpm). After washes and treatment with gentamicin to eliminate non-phagocytosed bacteria from the cells samples were (1) fixed and read by flow cytometry. (e) Gating of NLCs with FSC and SSC to eliminate cell debris and free bacteria from the analysis (left graph). Representative histogram describing the total population of NLCs, and composed of GFP negative cells and GFP positive cells (right graph). To assess phagocytosis, the percentage of GFP positive cells (f) and the mean fluorescence (g) were acquired. Data are presented as the mean values ± SEM. Results correspond to three pooled experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. (h) After washes and treatment with gentamicin, cells were also (2) lysed by Triton X-100, diluted in PBS and plated onto petri dishes. CFUs, corresponding to phagocytosed bacteria were counted in duplicates. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. To assess NLCs-dependent killing, P. aeruginosa PAO1 strain at a MOI of 2:1 was co-cultured with NLCs, treated with 10% NHS and CoMiX (15 μg/mL) for 1 h, plated on petri dishes to count the final bacterial CFUs in duplicates (i) . Conditions which were not co-cultured with NLCs were used as control (100% cell survival). Results are expressed as surviving bacteria compared to bacterial growth under the same conditions in the absence of NLCs. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01.
    M1 Macrophages Inos, supplied by Wanleibio, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m1 macrophages inos/product/Wanleibio
    Average 90 stars, based on 1 article reviews
    m1 macrophages inos - by Bioz Stars, 2025-12
    90/100 stars
    		  Buy from Supplier
    m1 macrophage npc interactions  (ATCC)
    98
    ATCC m1 macrophage npc interactions
    CoMiX-Fc enhances significantly phagocytosis of P. aeruginosa by PBMCs-derived <t>M1</t> <t>macrophages</t> and more importantly by Neutrophils-Like Cells, nevertheless, both CoMiX slightly improve NLCs-dependent antimicrobial activity against the bacteria. PMA-activated M1 macrophages, from four different healthy donors, were co-incubated with pHrodo-stained P. aeruginosa (PAO1) at a 12:1 bacteria-to-cell ratio with 10% NHS and in the presence or absence of 15 μg/mL CoMiX-Fc, CoMiX-FHR1 or CoMiX-irrelevant. Engulfment of bacteria was assessed after 30 min (left panels) and 1 h (right panels) by real-time Incucyte® microscope (a–c) . The percentage of pHrodo positive cells (a) and the intensity of pHrodo rationalised over the surface of cells and calculated as integrated intensity (b) were recorded. Data are presented as the mean values ± SEM. Results correspond to two independent experiments with macrophages from 4 healthy donors (three technical replicates per donor). Statistical analysis was performed using paired one-way ANOVA (to smooth inter-donor variability), followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01. (c) Representative incucyte images for the phagocytosis induced by serum and CoMiX-Fc over time. (d) Fluorescence microscopy image obtained on a wide field Axio Observer Z1, and treated on ImageJ of phagocytosed P. aeruginosa bacteria by PBMCs-derived M1 macrophages in presence of 10% serum and 15 μg/mL CoMiX-Fc. Red = wheat germ agglutinin Alexa-647, staining carbohydates residues of macrophages membranes; green = CellTrace™ CFSE stained P. aeruginosa bacteria. Scale bar = 25 μm. P. aeruginosa PAO1 strain and PAO1-GFP strain were co-cultured with NLCs at a 10:1 bacteria-to-cell ratio, treated with 2% NHS and CoMiX (15 μg/mL) and incubated for 20 min at 37 °C under agitation (200 rpm). After washes and treatment with gentamicin to eliminate non-phagocytosed bacteria from the cells samples were (1) fixed and read by flow cytometry. (e) Gating of NLCs with FSC and SSC to eliminate cell debris and free bacteria from the analysis (left graph). Representative histogram describing the total population of NLCs, and composed of GFP negative cells and GFP positive cells (right graph). To assess phagocytosis, the percentage of GFP positive cells (f) and the mean fluorescence (g) were acquired. Data are presented as the mean values ± SEM. Results correspond to three pooled experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. (h) After washes and treatment with gentamicin, cells were also (2) lysed by Triton X-100, diluted in PBS and plated onto petri dishes. CFUs, corresponding to phagocytosed bacteria were counted in duplicates. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. To assess NLCs-dependent killing, P. aeruginosa PAO1 strain at a MOI of 2:1 was co-cultured with NLCs, treated with 10% NHS and CoMiX (15 μg/mL) for 1 h, plated on petri dishes to count the final bacterial CFUs in duplicates (i) . Conditions which were not co-cultured with NLCs were used as control (100% cell survival). Results are expressed as surviving bacteria compared to bacterial growth under the same conditions in the absence of NLCs. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01.
    M1 Macrophage Npc Interactions, supplied by ATCC, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m1 macrophage npc interactions/product/ATCC
    Average 98 stars, based on 1 article reviews
    m1 macrophage npc interactions - by Bioz Stars, 2025-12
    98/100 stars
    		  Buy from Supplier
    gm csf  (PromoCell)
    94
    PromoCell gm csf
    CoMiX-Fc enhances significantly phagocytosis of P. aeruginosa by PBMCs-derived <t>M1</t> <t>macrophages</t> and more importantly by Neutrophils-Like Cells, nevertheless, both CoMiX slightly improve NLCs-dependent antimicrobial activity against the bacteria. PMA-activated M1 macrophages, from four different healthy donors, were co-incubated with pHrodo-stained P. aeruginosa (PAO1) at a 12:1 bacteria-to-cell ratio with 10% NHS and in the presence or absence of 15 μg/mL CoMiX-Fc, CoMiX-FHR1 or CoMiX-irrelevant. Engulfment of bacteria was assessed after 30 min (left panels) and 1 h (right panels) by real-time Incucyte® microscope (a–c) . The percentage of pHrodo positive cells (a) and the intensity of pHrodo rationalised over the surface of cells and calculated as integrated intensity (b) were recorded. Data are presented as the mean values ± SEM. Results correspond to two independent experiments with macrophages from 4 healthy donors (three technical replicates per donor). Statistical analysis was performed using paired one-way ANOVA (to smooth inter-donor variability), followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01. (c) Representative incucyte images for the phagocytosis induced by serum and CoMiX-Fc over time. (d) Fluorescence microscopy image obtained on a wide field Axio Observer Z1, and treated on ImageJ of phagocytosed P. aeruginosa bacteria by PBMCs-derived M1 macrophages in presence of 10% serum and 15 μg/mL CoMiX-Fc. Red = wheat germ agglutinin Alexa-647, staining carbohydates residues of macrophages membranes; green = CellTrace™ CFSE stained P. aeruginosa bacteria. Scale bar = 25 μm. P. aeruginosa PAO1 strain and PAO1-GFP strain were co-cultured with NLCs at a 10:1 bacteria-to-cell ratio, treated with 2% NHS and CoMiX (15 μg/mL) and incubated for 20 min at 37 °C under agitation (200 rpm). After washes and treatment with gentamicin to eliminate non-phagocytosed bacteria from the cells samples were (1) fixed and read by flow cytometry. (e) Gating of NLCs with FSC and SSC to eliminate cell debris and free bacteria from the analysis (left graph). Representative histogram describing the total population of NLCs, and composed of GFP negative cells and GFP positive cells (right graph). To assess phagocytosis, the percentage of GFP positive cells (f) and the mean fluorescence (g) were acquired. Data are presented as the mean values ± SEM. Results correspond to three pooled experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. (h) After washes and treatment with gentamicin, cells were also (2) lysed by Triton X-100, diluted in PBS and plated onto petri dishes. CFUs, corresponding to phagocytosed bacteria were counted in duplicates. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. To assess NLCs-dependent killing, P. aeruginosa PAO1 strain at a MOI of 2:1 was co-cultured with NLCs, treated with 10% NHS and CoMiX (15 μg/mL) for 1 h, plated on petri dishes to count the final bacterial CFUs in duplicates (i) . Conditions which were not co-cultured with NLCs were used as control (100% cell survival). Results are expressed as surviving bacteria compared to bacterial growth under the same conditions in the absence of NLCs. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01.
    Gm Csf, supplied by PromoCell, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gm csf/product/PromoCell
    Average 94 stars, based on 1 article reviews
    gm csf - by Bioz Stars, 2025-12
    94/100 stars
    		  Buy from Supplier
    m1 macrophages  (ATCC)
    98
    ATCC m1 macrophages
    Induction of efferocytosis and associated signaling in UVB-irradiated HCFs. (A) Real-time RT-PCR analysis for the efferocytotic ligand MFGE8 in control HCFs and HCFs irradiated with 150 mJ/cm 2 of UVB (BHCFs). n = 3. (B) Schematic diagram of the transwell-based co-culture system involving <t>M1</t> <t>macrophages</t> and either control HCFs or UVB (150 mJ/cm²)-irradiated HCFs (BHCFs). The transwell insert with 8 µm pores allowed M1 macrophages seeded in the upper chamber to migrate to the lower chamber and interact with apoptotic BHCFs. (C) Immunofluorescence analysis of in vitro efferocytosis in a co-culture of HCFs or BHCFs with TUNEL staining and PKH26 red-labeled M1 macrophages. Co-localization of apoptotic BHCFs and M1 macrophages ( white arrowheads ) was interpreted as evidence of efferocytosis. Scale bars : 50 µm. (D, E) Quantification of TUNEL-positive cells and efferocytic events as a proportion of total DAPI-positive cells in HCF or BHCF co-cultures. n = 3. Statistical analyses were performed using Student's t -test. ** P < 0.01 and * P < 0.05. Data are presented as mean ± SEM.
    M1 Macrophages, supplied by ATCC, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m1 macrophages/product/ATCC
    Average 98 stars, based on 1 article reviews
    m1 macrophages - by Bioz Stars, 2025-12
    98/100 stars
    		  Buy from Supplier
    involvement of m1/m2 macrophage polarization in reparative dentin formation  (Hasegawa Co Ltd)
    90
    Hasegawa Co Ltd involvement of m1/m2 macrophage polarization in reparative dentin formation
    Induction of efferocytosis and associated signaling in UVB-irradiated HCFs. (A) Real-time RT-PCR analysis for the efferocytotic ligand MFGE8 in control HCFs and HCFs irradiated with 150 mJ/cm 2 of UVB (BHCFs). n = 3. (B) Schematic diagram of the transwell-based co-culture system involving <t>M1</t> <t>macrophages</t> and either control HCFs or UVB (150 mJ/cm²)-irradiated HCFs (BHCFs). The transwell insert with 8 µm pores allowed M1 macrophages seeded in the upper chamber to migrate to the lower chamber and interact with apoptotic BHCFs. (C) Immunofluorescence analysis of in vitro efferocytosis in a co-culture of HCFs or BHCFs with TUNEL staining and PKH26 red-labeled M1 macrophages. Co-localization of apoptotic BHCFs and M1 macrophages ( white arrowheads ) was interpreted as evidence of efferocytosis. Scale bars : 50 µm. (D, E) Quantification of TUNEL-positive cells and efferocytic events as a proportion of total DAPI-positive cells in HCF or BHCF co-cultures. n = 3. Statistical analyses were performed using Student's t -test. ** P < 0.01 and * P < 0.05. Data are presented as mean ± SEM.
    Involvement Of M1/M2 Macrophage Polarization In Reparative Dentin Formation, supplied by Hasegawa Co Ltd, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/involvement of m1/m2 macrophage polarization in reparative dentin formation/product/Hasegawa Co Ltd
    Average 90 stars, based on 1 article reviews
    involvement of m1/m2 macrophage polarization in reparative dentin formation - by Bioz Stars, 2025-12
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    CoMiX-Fc enhances significantly phagocytosis of P. aeruginosa by PBMCs-derived M1 macrophages and more importantly by Neutrophils-Like Cells, nevertheless, both CoMiX slightly improve NLCs-dependent antimicrobial activity against the bacteria. PMA-activated M1 macrophages, from four different healthy donors, were co-incubated with pHrodo-stained P. aeruginosa (PAO1) at a 12:1 bacteria-to-cell ratio with 10% NHS and in the presence or absence of 15 μg/mL CoMiX-Fc, CoMiX-FHR1 or CoMiX-irrelevant. Engulfment of bacteria was assessed after 30 min (left panels) and 1 h (right panels) by real-time Incucyte® microscope (a–c) . The percentage of pHrodo positive cells (a) and the intensity of pHrodo rationalised over the surface of cells and calculated as integrated intensity (b) were recorded. Data are presented as the mean values ± SEM. Results correspond to two independent experiments with macrophages from 4 healthy donors (three technical replicates per donor). Statistical analysis was performed using paired one-way ANOVA (to smooth inter-donor variability), followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01. (c) Representative incucyte images for the phagocytosis induced by serum and CoMiX-Fc over time. (d) Fluorescence microscopy image obtained on a wide field Axio Observer Z1, and treated on ImageJ of phagocytosed P. aeruginosa bacteria by PBMCs-derived M1 macrophages in presence of 10% serum and 15 μg/mL CoMiX-Fc. Red = wheat germ agglutinin Alexa-647, staining carbohydates residues of macrophages membranes; green = CellTrace™ CFSE stained P. aeruginosa bacteria. Scale bar = 25 μm. P. aeruginosa PAO1 strain and PAO1-GFP strain were co-cultured with NLCs at a 10:1 bacteria-to-cell ratio, treated with 2% NHS and CoMiX (15 μg/mL) and incubated for 20 min at 37 °C under agitation (200 rpm). After washes and treatment with gentamicin to eliminate non-phagocytosed bacteria from the cells samples were (1) fixed and read by flow cytometry. (e) Gating of NLCs with FSC and SSC to eliminate cell debris and free bacteria from the analysis (left graph). Representative histogram describing the total population of NLCs, and composed of GFP negative cells and GFP positive cells (right graph). To assess phagocytosis, the percentage of GFP positive cells (f) and the mean fluorescence (g) were acquired. Data are presented as the mean values ± SEM. Results correspond to three pooled experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. (h) After washes and treatment with gentamicin, cells were also (2) lysed by Triton X-100, diluted in PBS and plated onto petri dishes. CFUs, corresponding to phagocytosed bacteria were counted in duplicates. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. To assess NLCs-dependent killing, P. aeruginosa PAO1 strain at a MOI of 2:1 was co-cultured with NLCs, treated with 10% NHS and CoMiX (15 μg/mL) for 1 h, plated on petri dishes to count the final bacterial CFUs in duplicates (i) . Conditions which were not co-cultured with NLCs were used as control (100% cell survival). Results are expressed as surviving bacteria compared to bacterial growth under the same conditions in the absence of NLCs. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01.

    Journal: eBioMedicine

    Article Title: Directed-complement killing of Pseudomonas aeruginosa protects against lethal pneumonia

    doi: 10.1016/j.ebiom.2025.105926

    Figure Lengend Snippet: CoMiX-Fc enhances significantly phagocytosis of P. aeruginosa by PBMCs-derived M1 macrophages and more importantly by Neutrophils-Like Cells, nevertheless, both CoMiX slightly improve NLCs-dependent antimicrobial activity against the bacteria. PMA-activated M1 macrophages, from four different healthy donors, were co-incubated with pHrodo-stained P. aeruginosa (PAO1) at a 12:1 bacteria-to-cell ratio with 10% NHS and in the presence or absence of 15 μg/mL CoMiX-Fc, CoMiX-FHR1 or CoMiX-irrelevant. Engulfment of bacteria was assessed after 30 min (left panels) and 1 h (right panels) by real-time Incucyte® microscope (a–c) . The percentage of pHrodo positive cells (a) and the intensity of pHrodo rationalised over the surface of cells and calculated as integrated intensity (b) were recorded. Data are presented as the mean values ± SEM. Results correspond to two independent experiments with macrophages from 4 healthy donors (three technical replicates per donor). Statistical analysis was performed using paired one-way ANOVA (to smooth inter-donor variability), followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01. (c) Representative incucyte images for the phagocytosis induced by serum and CoMiX-Fc over time. (d) Fluorescence microscopy image obtained on a wide field Axio Observer Z1, and treated on ImageJ of phagocytosed P. aeruginosa bacteria by PBMCs-derived M1 macrophages in presence of 10% serum and 15 μg/mL CoMiX-Fc. Red = wheat germ agglutinin Alexa-647, staining carbohydates residues of macrophages membranes; green = CellTrace™ CFSE stained P. aeruginosa bacteria. Scale bar = 25 μm. P. aeruginosa PAO1 strain and PAO1-GFP strain were co-cultured with NLCs at a 10:1 bacteria-to-cell ratio, treated with 2% NHS and CoMiX (15 μg/mL) and incubated for 20 min at 37 °C under agitation (200 rpm). After washes and treatment with gentamicin to eliminate non-phagocytosed bacteria from the cells samples were (1) fixed and read by flow cytometry. (e) Gating of NLCs with FSC and SSC to eliminate cell debris and free bacteria from the analysis (left graph). Representative histogram describing the total population of NLCs, and composed of GFP negative cells and GFP positive cells (right graph). To assess phagocytosis, the percentage of GFP positive cells (f) and the mean fluorescence (g) were acquired. Data are presented as the mean values ± SEM. Results correspond to three pooled experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. (h) After washes and treatment with gentamicin, cells were also (2) lysed by Triton X-100, diluted in PBS and plated onto petri dishes. CFUs, corresponding to phagocytosed bacteria were counted in duplicates. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗∗∗∗p < 0.0001. To assess NLCs-dependent killing, P. aeruginosa PAO1 strain at a MOI of 2:1 was co-cultured with NLCs, treated with 10% NHS and CoMiX (15 μg/mL) for 1 h, plated on petri dishes to count the final bacterial CFUs in duplicates (i) . Conditions which were not co-cultured with NLCs were used as control (100% cell survival). Results are expressed as surviving bacteria compared to bacterial growth under the same conditions in the absence of NLCs. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments (2 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: ∗p < 0.05; ∗∗p < 0.01.

    Article Snippet: Peripheral blood mononuclear cells (PBMCs) from healthy donors (Luxembourg Red Cross, MAN_SCE_24_008) were isolated from buffy coats and differentiated into M1 macrophages using and following the manufacturer's instructions of PromoCell macrophage generation medium (PromoCell, Germany).

    Techniques: Derivative Assay, Activity Assay, Bacteria, Incubation, Staining, Microscopy, Fluorescence, Cell Culture, Flow Cytometry, Control

    Induction of efferocytosis and associated signaling in UVB-irradiated HCFs. (A) Real-time RT-PCR analysis for the efferocytotic ligand MFGE8 in control HCFs and HCFs irradiated with 150 mJ/cm 2 of UVB (BHCFs). n = 3. (B) Schematic diagram of the transwell-based co-culture system involving M1 macrophages and either control HCFs or UVB (150 mJ/cm²)-irradiated HCFs (BHCFs). The transwell insert with 8 µm pores allowed M1 macrophages seeded in the upper chamber to migrate to the lower chamber and interact with apoptotic BHCFs. (C) Immunofluorescence analysis of in vitro efferocytosis in a co-culture of HCFs or BHCFs with TUNEL staining and PKH26 red-labeled M1 macrophages. Co-localization of apoptotic BHCFs and M1 macrophages ( white arrowheads ) was interpreted as evidence of efferocytosis. Scale bars : 50 µm. (D, E) Quantification of TUNEL-positive cells and efferocytic events as a proportion of total DAPI-positive cells in HCF or BHCF co-cultures. n = 3. Statistical analyses were performed using Student's t -test. ** P < 0.01 and * P < 0.05. Data are presented as mean ± SEM.

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Efferocytosis by Macrophages Attenuates Inflammatory Responses Following Ultraviolet B-Induced Apoptosis in Corneal Stromal Cells

    doi: 10.1167/iovs.66.9.49

    Figure Lengend Snippet: Induction of efferocytosis and associated signaling in UVB-irradiated HCFs. (A) Real-time RT-PCR analysis for the efferocytotic ligand MFGE8 in control HCFs and HCFs irradiated with 150 mJ/cm 2 of UVB (BHCFs). n = 3. (B) Schematic diagram of the transwell-based co-culture system involving M1 macrophages and either control HCFs or UVB (150 mJ/cm²)-irradiated HCFs (BHCFs). The transwell insert with 8 µm pores allowed M1 macrophages seeded in the upper chamber to migrate to the lower chamber and interact with apoptotic BHCFs. (C) Immunofluorescence analysis of in vitro efferocytosis in a co-culture of HCFs or BHCFs with TUNEL staining and PKH26 red-labeled M1 macrophages. Co-localization of apoptotic BHCFs and M1 macrophages ( white arrowheads ) was interpreted as evidence of efferocytosis. Scale bars : 50 µm. (D, E) Quantification of TUNEL-positive cells and efferocytic events as a proportion of total DAPI-positive cells in HCF or BHCF co-cultures. n = 3. Statistical analyses were performed using Student's t -test. ** P < 0.01 and * P < 0.05. Data are presented as mean ± SEM.

    Article Snippet: M1 macrophages were derived from the THP-1 cell line (TIB-202; ATCC, Manassas, VA, USA) following established methods.

    Techniques: Irradiation, Quantitative RT-PCR, Control, Co-Culture Assay, Immunofluorescence, In Vitro, TUNEL Assay, Staining, Labeling

    Efferocytosis and inflammation-related signals in UVB-irradiated HCFs co-cultured with M1 macrophages within an in vitro efferocytosis model. (A) Schematic diagram of the transwell-based co-culture system involving M1 macrophages and either control HCFs or UVB (150 mJ/cm²)-irradiated HCFs (BHCFs). (B, C) Real-time RT-PCR analysis of the efferocytosis-related ligand MFGE8 and the receptor MERTK in three groups: non-irradiated HCFs without co-culture with M1 macrophages (i.e., HCF only), non-irradiated HCFs co-cultured with M1 macrophages (i.e., HCF and M1), and UVB-irradiated HCFs co-cultured with M1 macrophages (i.e., BHCF and M1). The cells used for analysis were confined to the bottom area of the well (inside a dotted rounded rectangle ). n = 3. (D) Representative Western blot images of MFG-E8 and MERTK in three groups. (E, F) Real-time RT-PCR analysis of the innate immune-related inflammatory cytokines IL1B and IL6 in three groups. n = 3. ( G ) ELISA analysis of IL-6 protein levels in the culture supernatant from three groups. n = 3. Statistical analyses were performed using one-way ANOVA followed by Tukey's post-hoc test. **** P < 0.0001, *** P < 0.001, ** P < 0.01, and * P < 0.05; ns, not significant. Data are presented as mean ± SEM.

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Efferocytosis by Macrophages Attenuates Inflammatory Responses Following Ultraviolet B-Induced Apoptosis in Corneal Stromal Cells

    doi: 10.1167/iovs.66.9.49

    Figure Lengend Snippet: Efferocytosis and inflammation-related signals in UVB-irradiated HCFs co-cultured with M1 macrophages within an in vitro efferocytosis model. (A) Schematic diagram of the transwell-based co-culture system involving M1 macrophages and either control HCFs or UVB (150 mJ/cm²)-irradiated HCFs (BHCFs). (B, C) Real-time RT-PCR analysis of the efferocytosis-related ligand MFGE8 and the receptor MERTK in three groups: non-irradiated HCFs without co-culture with M1 macrophages (i.e., HCF only), non-irradiated HCFs co-cultured with M1 macrophages (i.e., HCF and M1), and UVB-irradiated HCFs co-cultured with M1 macrophages (i.e., BHCF and M1). The cells used for analysis were confined to the bottom area of the well (inside a dotted rounded rectangle ). n = 3. (D) Representative Western blot images of MFG-E8 and MERTK in three groups. (E, F) Real-time RT-PCR analysis of the innate immune-related inflammatory cytokines IL1B and IL6 in three groups. n = 3. ( G ) ELISA analysis of IL-6 protein levels in the culture supernatant from three groups. n = 3. Statistical analyses were performed using one-way ANOVA followed by Tukey's post-hoc test. **** P < 0.0001, *** P < 0.001, ** P < 0.01, and * P < 0.05; ns, not significant. Data are presented as mean ± SEM.

    Article Snippet: M1 macrophages were derived from the THP-1 cell line (TIB-202; ATCC, Manassas, VA, USA) following established methods.

    Techniques: Irradiation, Cell Culture, In Vitro, Co-Culture Assay, Control, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay

    Paracrine regulation of microenvironmental M1 macrophages in response to the efferocytosis of UVB-irradiated HCFs. (A) Schematic diagram of the transwell-based co-culture system involving M1 macrophages and either control HCFs or UVB (150 mJ/cm²)-irradiated HCFs (BHCFs). (B–D) Real-time RT-PCR analysis of the pro-inflammatory cytokines IL1B , IL6 , and the anti-inflammatory cytokine TGFB1 , in microenvironmental M1 macrophages under three co-culture conditions: M1 macrophages without co-culture with HCFs or BHCFs (i.e., M1 only), non-irradiated HCFs co-cultured with M1 macrophages (i.e., HCF and M1), and BHCFs co-cultured with M1 macrophages (i.e., BHCF and M1). The cells used for analysis were limited to the microenvironmental M1 macrophages on the transwell insert (inside a dotted rounded rectangle). n = 3. (E) ELISA analysis of IL-6 protein levels in the supernatants from three groups. n = 3. (F–J) Real-time RT-PCR analysis of the monocyte/myeloid/M1 macrophage-related markers, including CD14 , ITGAM (i.e., CD11B ), ITGAX (i.e., CD11C ), CD68 , and CD80 in three groups. n = 3. (K) Representative Western blot images of CD80 in three groups. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. **** P < 0.0001, *** P < 0.001, ** P < 0.01, and * P < 0.05; ns, not significant. Data are presented as mean ± SEM.

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Efferocytosis by Macrophages Attenuates Inflammatory Responses Following Ultraviolet B-Induced Apoptosis in Corneal Stromal Cells

    doi: 10.1167/iovs.66.9.49

    Figure Lengend Snippet: Paracrine regulation of microenvironmental M1 macrophages in response to the efferocytosis of UVB-irradiated HCFs. (A) Schematic diagram of the transwell-based co-culture system involving M1 macrophages and either control HCFs or UVB (150 mJ/cm²)-irradiated HCFs (BHCFs). (B–D) Real-time RT-PCR analysis of the pro-inflammatory cytokines IL1B , IL6 , and the anti-inflammatory cytokine TGFB1 , in microenvironmental M1 macrophages under three co-culture conditions: M1 macrophages without co-culture with HCFs or BHCFs (i.e., M1 only), non-irradiated HCFs co-cultured with M1 macrophages (i.e., HCF and M1), and BHCFs co-cultured with M1 macrophages (i.e., BHCF and M1). The cells used for analysis were limited to the microenvironmental M1 macrophages on the transwell insert (inside a dotted rounded rectangle). n = 3. (E) ELISA analysis of IL-6 protein levels in the supernatants from three groups. n = 3. (F–J) Real-time RT-PCR analysis of the monocyte/myeloid/M1 macrophage-related markers, including CD14 , ITGAM (i.e., CD11B ), ITGAX (i.e., CD11C ), CD68 , and CD80 in three groups. n = 3. (K) Representative Western blot images of CD80 in three groups. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. **** P < 0.0001, *** P < 0.001, ** P < 0.01, and * P < 0.05; ns, not significant. Data are presented as mean ± SEM.

    Article Snippet: M1 macrophages were derived from the THP-1 cell line (TIB-202; ATCC, Manassas, VA, USA) following established methods.

    Techniques: Irradiation, Co-Culture Assay, Control, Quantitative RT-PCR, Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot