Journal: International Journal of Molecular Sciences

Article Title: Mesenchymal Stem Cell Exosomes as Immunomodulatory Therapy for Corneal Scarring

doi: 10.3390/ijms24087456

Figure Lengend Snippet: Macrophage polarization in laser-injured rat corneas after 5 days of MSC-exo or PBS topical application. Fold change of gene expression of M1 phenotype, CD80 ( A ), CD86 ( B ), and NOS2 ( C ), and M2 phenotype, CD163 ( D ), CD206 ( E ), and ARG1 ( F ) in the injured corneas from the uninjured corneas. ( G ) M2/M1 gene ratio was significantly higher in the corneas treated with MSC-exo. ( H ) Corresponding immunohistochemistry of M1 and M2 polarized macrophage markers was performed and viewed at the laser-injury site. The images were orientated such that the corneal epithelium (Ep) appeared at the top, followed by the stroma (S), and the endothelium (En) at the bottom. The cells expressing M1 polarized macrophage markers, CD80 ( I ) and CD86 ( J ), were counted and compared to those expressing M2 macrophage markers, CD163 ( K ) and CD206 ( L ). ( M ) The M2/M1 positive-cells ratio was significantly higher in the corneas treated with MSC-exo than in the PBS group. * p < 0.05. Scale bars = 50 µm.

Article Snippet: Samples were treated with ice-cold 50 mM ammonium chloride (Sigma-Aldrich), saponin-permeabilized, and blocked with 2% BSA and 5% NGS, followed by incubation with primary antibodies: Mouse monoclonal anti-fibronectin (Sigma-Aldrich), CD31 (Thermo Fisher), CD80 (Lifespan Biosciences, Seattle, WA, USA), CD163 (Bio-Rad Laboratories) and α-SMA (Agilent) antibodies, and rabbit polyclonal anti-collagen 3A1 antibody (Novus Biologicals, Littleton, CO, USA), LYVE-1 (Abcam, Cambridge, UK), CD86 (Abcam) and CD206 (Abcam) antibodies for 2 h at room temperature.

Techniques: Expressing, Immunohistochemistry

Journal: International Journal of Molecular Sciences

Article Title: Mesenchymal Stem Cell Exosomes as Immunomodulatory Therapy for Corneal Scarring

doi: 10.3390/ijms24087456

Figure Lengend Snippet: Primer sequence used in the RT-PCR analysis.

Article Snippet: Samples were treated with ice-cold 50 mM ammonium chloride (Sigma-Aldrich), saponin-permeabilized, and blocked with 2% BSA and 5% NGS, followed by incubation with primary antibodies: Mouse monoclonal anti-fibronectin (Sigma-Aldrich), CD31 (Thermo Fisher), CD80 (Lifespan Biosciences, Seattle, WA, USA), CD163 (Bio-Rad Laboratories) and α-SMA (Agilent) antibodies, and rabbit polyclonal anti-collagen 3A1 antibody (Novus Biologicals, Littleton, CO, USA), LYVE-1 (Abcam, Cambridge, UK), CD86 (Abcam) and CD206 (Abcam) antibodies for 2 h at room temperature.

Techniques: Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Mesenchymal Stem Cell Exosomes as Immunomodulatory Therapy for Corneal Scarring

doi: 10.3390/ijms24087456

Figure Lengend Snippet: Macrophage polarization in laser-injured rat corneas after 5 days of MSC-exo or PBS topical application. Fold change of gene expression of M1 phenotype, CD80 ( A ), CD86 ( B ), and NOS2 ( C ), and M2 phenotype, CD163 ( D ), CD206 ( E ), and ARG1 ( F ) in the injured corneas from the uninjured corneas. ( G ) M2/M1 gene ratio was significantly higher in the corneas treated with MSC-exo. ( H ) Corresponding immunohistochemistry of M1 and M2 polarized macrophage markers was performed and viewed at the laser-injury site. The images were orientated such that the corneal epithelium (Ep) appeared at the top, followed by the stroma (S), and the endothelium (En) at the bottom. The cells expressing M1 polarized macrophage markers, CD80 ( I ) and CD86 ( J ), were counted and compared to those expressing M2 macrophage markers, CD163 ( K ) and CD206 ( L ). ( M ) The M2/M1 positive-cells ratio was significantly higher in the corneas treated with MSC-exo than in the PBS group. * p < 0.05. Scale bars = 50 µm.

Article Snippet: Samples were treated with ice-cold 50 mM ammonium chloride (Sigma-Aldrich), saponin-permeabilized, and blocked with 2% BSA and 5% NGS, followed by incubation with primary antibodies: Mouse monoclonal anti-fibronectin (Sigma-Aldrich), CD31 (Thermo Fisher), CD80 (Lifespan Biosciences, Seattle, WA, USA), CD163 (Bio-Rad Laboratories) and α-SMA (Agilent) antibodies, and rabbit polyclonal anti-collagen 3A1 antibody (Novus Biologicals, Littleton, CO, USA), LYVE-1 (Abcam, Cambridge, UK), CD86 (Abcam) and CD206 (Abcam) antibodies for 2 h at room temperature.

Techniques: Expressing, Immunohistochemistry

Journal: International Journal of Molecular Sciences

Article Title: Mesenchymal Stem Cell Exosomes as Immunomodulatory Therapy for Corneal Scarring

doi: 10.3390/ijms24087456

Figure Lengend Snippet: Primer sequence used in the RT-PCR analysis.

Article Snippet: Samples were treated with ice-cold 50 mM ammonium chloride (Sigma-Aldrich), saponin-permeabilized, and blocked with 2% BSA and 5% NGS, followed by incubation with primary antibodies: Mouse monoclonal anti-fibronectin (Sigma-Aldrich), CD31 (Thermo Fisher), CD80 (Lifespan Biosciences, Seattle, WA, USA), CD163 (Bio-Rad Laboratories) and α-SMA (Agilent) antibodies, and rabbit polyclonal anti-collagen 3A1 antibody (Novus Biologicals, Littleton, CO, USA), LYVE-1 (Abcam, Cambridge, UK), CD86 (Abcam) and CD206 (Abcam) antibodies for 2 h at room temperature.

Techniques: Sequencing