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    Thermo Fisher rat msc conditioned media
    <t>MSC-CM</t> suppressed the inflammatory reaction and enhanced phagocytosis in LPS-stimulated microglia. (A): To determine the effect of MSC-CM on the level of cytokines in LPS-stimulated microglia, we applied MSC-CM for 24, 48, and 72 hours to LPS-stimulated microglia. LPS increased mRNA expression of TNF-α, IL-1β, IL-6, and iNOS, and nitrate secretion in LPS-stimulated microglia, whereas MSC-CM inhibited the inflammatory reaction. (B): Immunostaining shows that MSC-CM significantly suppressed iNOS expression in LPS-stimulated microglia. The microglia and nuclei were stained with Iba-1 and DAPI, respectively. (C): LPS-stimulated microglia ingested more latex beads than did untreated controls, and the phagocytic activity of microglia was significantly enhanced when the MSC-CM was applied to LPS-stimulated microglia following LPS treatment. The data are means ± SEM of three independent experiments. ∗, p < .05; ∗∗, p < .01; ∗∗∗, p < .001; in comparison with control. §, p < .05; §§, p < .05; §§§, p < .001; in comparison with LPS. Abbreviations: CON, control; DAPI, 4′,6-diamidino-2-phenylindole; hr, hour; iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal <t>cell-conditioned</t> media; RQ, relative quantity; TNF, tumor necrosis factor.
    Rat Msc Conditioned Media, supplied by Thermo Fisher, 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/product/rat+msc+conditioned+media/pmc05070497-81-0-18?v=Thermo+Fisher
    Average 90 stars, based on 1 article reviews
    rat msc conditioned media - by Bioz Stars, 2026-08
    90/100 stars

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    1) Product Images from "Mesenchymal Stem Cells Modulate the Functional Properties of Microglia via TGF-β Secretion"

    Article Title: Mesenchymal Stem Cells Modulate the Functional Properties of Microglia via TGF-β Secretion

    Journal: Stem Cells Translational Medicine

    doi: 10.5966/sctm.2015-0217

    MSC-CM suppressed the inflammatory reaction and enhanced phagocytosis in LPS-stimulated microglia. (A): To determine the effect of MSC-CM on the level of cytokines in LPS-stimulated microglia, we applied MSC-CM for 24, 48, and 72 hours to LPS-stimulated microglia. LPS increased mRNA expression of TNF-α, IL-1β, IL-6, and iNOS, and nitrate secretion in LPS-stimulated microglia, whereas MSC-CM inhibited the inflammatory reaction. (B): Immunostaining shows that MSC-CM significantly suppressed iNOS expression in LPS-stimulated microglia. The microglia and nuclei were stained with Iba-1 and DAPI, respectively. (C): LPS-stimulated microglia ingested more latex beads than did untreated controls, and the phagocytic activity of microglia was significantly enhanced when the MSC-CM was applied to LPS-stimulated microglia following LPS treatment. The data are means ± SEM of three independent experiments. ∗, p < .05; ∗∗, p < .01; ∗∗∗, p < .001; in comparison with control. §, p < .05; §§, p < .05; §§§, p < .001; in comparison with LPS. Abbreviations: CON, control; DAPI, 4′,6-diamidino-2-phenylindole; hr, hour; iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; RQ, relative quantity; TNF, tumor necrosis factor.
    Figure Legend Snippet: MSC-CM suppressed the inflammatory reaction and enhanced phagocytosis in LPS-stimulated microglia. (A): To determine the effect of MSC-CM on the level of cytokines in LPS-stimulated microglia, we applied MSC-CM for 24, 48, and 72 hours to LPS-stimulated microglia. LPS increased mRNA expression of TNF-α, IL-1β, IL-6, and iNOS, and nitrate secretion in LPS-stimulated microglia, whereas MSC-CM inhibited the inflammatory reaction. (B): Immunostaining shows that MSC-CM significantly suppressed iNOS expression in LPS-stimulated microglia. The microglia and nuclei were stained with Iba-1 and DAPI, respectively. (C): LPS-stimulated microglia ingested more latex beads than did untreated controls, and the phagocytic activity of microglia was significantly enhanced when the MSC-CM was applied to LPS-stimulated microglia following LPS treatment. The data are means ± SEM of three independent experiments. ∗, p < .05; ∗∗, p < .01; ∗∗∗, p < .001; in comparison with control. §, p < .05; §§, p < .05; §§§, p < .001; in comparison with LPS. Abbreviations: CON, control; DAPI, 4′,6-diamidino-2-phenylindole; hr, hour; iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; RQ, relative quantity; TNF, tumor necrosis factor.

    Techniques Used: Expressing, Immunostaining, Staining, Activity Assay, Comparison, Control

    MSC-CM restored CX3CR1, CD206, and CD200R expression in LPS-stimulated microglia. To investigate the effect of MSC-CM on the expression of CD86, CX3CR1, CD206, and CD200R in LPS-stimulated microglia, we performed flow cytometry. (A): Microglia were treated with LPS for 24 hours and MSC-CM was incubated for 72 hours. The isotype control was indicated as gray color. Control group (red), LPS group (green), and LPS + MSC-CM (blue) were indicated, respectively. The table box beside the diagram indicates the percentage of positive cells in each group. (B): Rather than divide this microglia population further by imposing strict boundaries on a continuous expression pattern, we then determined MFI of the entire remaining population for each marker, and we calculated the M2:M1 MFI ratio (CX3CR1, CD206, or CD200R/CD86). MSC-CM restored CX3CR1, CD206, and CD200R MFI, whereas it inhibited the CD86 in LPS-stimulated microglia, showing the normalization of M2:M1 MFI ratio. (C): Immunofluorescence results showed a similar pattern with flow cytometry analysis. Microglia was stained with Iba-1 and DAPI, respectively. (D): MSC-CM restored mRNA expression of CX3CR1 and CD206 in LPS-stimulated microglia following to incubation time. The data are means ± SEM of three independent experiments. §, p < .001 in comparison with control (CON); §§, p < .01; §§§, p < .001 in comparison with LPS. Abbreviations: CON, control; DAPI, 4′,6-diamidino-2-phenylindole; hr, hour; LPS, lipopolysaccharide; MFI, mean fluorescence intensity; MSC-CM, mesenchymal stromal cell-conditioned media; RQ, relative quantity.
    Figure Legend Snippet: MSC-CM restored CX3CR1, CD206, and CD200R expression in LPS-stimulated microglia. To investigate the effect of MSC-CM on the expression of CD86, CX3CR1, CD206, and CD200R in LPS-stimulated microglia, we performed flow cytometry. (A): Microglia were treated with LPS for 24 hours and MSC-CM was incubated for 72 hours. The isotype control was indicated as gray color. Control group (red), LPS group (green), and LPS + MSC-CM (blue) were indicated, respectively. The table box beside the diagram indicates the percentage of positive cells in each group. (B): Rather than divide this microglia population further by imposing strict boundaries on a continuous expression pattern, we then determined MFI of the entire remaining population for each marker, and we calculated the M2:M1 MFI ratio (CX3CR1, CD206, or CD200R/CD86). MSC-CM restored CX3CR1, CD206, and CD200R MFI, whereas it inhibited the CD86 in LPS-stimulated microglia, showing the normalization of M2:M1 MFI ratio. (C): Immunofluorescence results showed a similar pattern with flow cytometry analysis. Microglia was stained with Iba-1 and DAPI, respectively. (D): MSC-CM restored mRNA expression of CX3CR1 and CD206 in LPS-stimulated microglia following to incubation time. The data are means ± SEM of three independent experiments. §, p < .001 in comparison with control (CON); §§, p < .01; §§§, p < .001 in comparison with LPS. Abbreviations: CON, control; DAPI, 4′,6-diamidino-2-phenylindole; hr, hour; LPS, lipopolysaccharide; MFI, mean fluorescence intensity; MSC-CM, mesenchymal stromal cell-conditioned media; RQ, relative quantity.

    Techniques Used: Expressing, Flow Cytometry, Incubation, Control, Marker, Immunofluorescence, Staining, Comparison, Fluorescence

    TGF-β mediates the effect of MSC-CM on LPS-stimulated microglia. To determine which soluble factor was involved in the effect of MSC-CM on LPS-stimulated microglia, we treated LPS-stimulated microglia with MSC-CM + a TGF-βR inhibitor (A), a CX3CL1 antibody (B), or siMSC-CM (siMSC-CM indicates conditioned medium that was obtained from TGF-β siRNA-transfected MSCs) (C). LPS increased TNF-α, IL-1β, IL-6, and iNOS mRNA levels, and MSC-CM inhibited the increased cytokines and led to CX3CR1 restoration. TGF-β inhibition abolished the effect of MSC-CM on LPS-stimulated microglia, whereas CX3CL1 antibody did not. siMSC-CM did not affect LPS-stimulated microglia. The data are means ± SEM of three independent experiments. ∗, p < .01; ∗∗, p < .001; in comparison with control (CON). §, p < .05; §§, p < .01; §§§, p < .001; in comparison with LPS. †, p < .05; ††, p < .01; †††, p < .001; in comparison with two groups (LPS + MSC-CM vs. LPS + MSC-CM + antibody/inhibitor or siMSC-CM). Abbreviations: CON, control; iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; n.s., not significant; RQ, relative quantity; TGF, transforming growth factor; TNF, tumor necrosis factor.
    Figure Legend Snippet: TGF-β mediates the effect of MSC-CM on LPS-stimulated microglia. To determine which soluble factor was involved in the effect of MSC-CM on LPS-stimulated microglia, we treated LPS-stimulated microglia with MSC-CM + a TGF-βR inhibitor (A), a CX3CL1 antibody (B), or siMSC-CM (siMSC-CM indicates conditioned medium that was obtained from TGF-β siRNA-transfected MSCs) (C). LPS increased TNF-α, IL-1β, IL-6, and iNOS mRNA levels, and MSC-CM inhibited the increased cytokines and led to CX3CR1 restoration. TGF-β inhibition abolished the effect of MSC-CM on LPS-stimulated microglia, whereas CX3CL1 antibody did not. siMSC-CM did not affect LPS-stimulated microglia. The data are means ± SEM of three independent experiments. ∗, p < .01; ∗∗, p < .001; in comparison with control (CON). §, p < .05; §§, p < .01; §§§, p < .001; in comparison with LPS. †, p < .05; ††, p < .01; †††, p < .001; in comparison with two groups (LPS + MSC-CM vs. LPS + MSC-CM + antibody/inhibitor or siMSC-CM). Abbreviations: CON, control; iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; n.s., not significant; RQ, relative quantity; TGF, transforming growth factor; TNF, tumor necrosis factor.

    Techniques Used: Transfection, Inhibition, Comparison, Control

    TGF-βR inhibition alone can abolish the effect of MSC-CM in LPS-stimulated microglia. To investigate whether CX3CL1 plays a role in the anti-inflammatory effect of TGF-β in MSC-CM, we added a CX3CL1 antibody, a TGF-βR inhibitor, and both together to MSC-CM. LPS reduced CX3CR1 (A) and increased TNF-α mRNA expression (B), IL-1β mRNA expression (C), IL-6 mRNA expression (D), and iNOS mRNA expression (E), and. MSC-CM rescued these changes. The TGF-βR inhibitor alone showed the same effect on LPS-stimulated microglia, regardless of the presence of the CX3CL1 antibody. When the TGF-βR inhibitor was added to the MSC-CM, the effect of the MSC-CM on TNF-α, IL-1β, IL-6, iNOS, and CX3CR1 was abolished. The data are means ± SEM of three independent experiments. ∗, p < .05; ∗∗, p < .01; ∗∗∗, p < .001; in comparison with control. §, p < .05; §§, p < .01; §§§, p < .001; in comparison with LPS. †, p < .05; †††, p < .001; compared between two groups. Abbreviations: iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; RQ, relative quantity; TGF, transforming growth factor; TNF, tumor necrosis factor.
    Figure Legend Snippet: TGF-βR inhibition alone can abolish the effect of MSC-CM in LPS-stimulated microglia. To investigate whether CX3CL1 plays a role in the anti-inflammatory effect of TGF-β in MSC-CM, we added a CX3CL1 antibody, a TGF-βR inhibitor, and both together to MSC-CM. LPS reduced CX3CR1 (A) and increased TNF-α mRNA expression (B), IL-1β mRNA expression (C), IL-6 mRNA expression (D), and iNOS mRNA expression (E), and. MSC-CM rescued these changes. The TGF-βR inhibitor alone showed the same effect on LPS-stimulated microglia, regardless of the presence of the CX3CL1 antibody. When the TGF-βR inhibitor was added to the MSC-CM, the effect of the MSC-CM on TNF-α, IL-1β, IL-6, iNOS, and CX3CR1 was abolished. The data are means ± SEM of three independent experiments. ∗, p < .05; ∗∗, p < .01; ∗∗∗, p < .001; in comparison with control. §, p < .05; §§, p < .01; §§§, p < .001; in comparison with LPS. †, p < .05; †††, p < .001; compared between two groups. Abbreviations: iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; RQ, relative quantity; TGF, transforming growth factor; TNF, tumor necrosis factor.

    Techniques Used: Inhibition, Expressing, Comparison, Control

    Recombinant TGF-β showed an effect similar to that of MSC-CM on LPS-stimulated microglia. Recombinant TGF-β (rTGF-β, 10 ng/ml) induced similar effects as did MSC-CM. On the basis of a previous study, recombinant CX3CL1 (rCX3CL1, 10 ng/ml) was also tested. The rTGF-β and rCX3CL1 were applied for 24 hours. rTGF-β rescued the reduced CX3CR1 expression (A) and inhibited the increased gene expression of TNF-α (B), IL-1β (C), IL-6 (D), and iNOS (E), similar to MSC-CM in LPS-stimulated microglia. rCX3CL1 (10 ng/ml) also inhibited IL-1β and iNOS expression and restored CX3CR1 expression but did not affect TNF-α and IL-6 expression. The data are means ± SEM of three independent experiments. ∗, p < .05; ∗∗∗, p < .001; in comparison with control. §, p < .05; §§§, p < .001; in comparison with LPS. ††, p < .05; †††, p < .001; compared between two groups. Abbreviations: iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; RQ, relative quantity; TGF, transforming growth factor; TNF, tumor necrosis factor.
    Figure Legend Snippet: Recombinant TGF-β showed an effect similar to that of MSC-CM on LPS-stimulated microglia. Recombinant TGF-β (rTGF-β, 10 ng/ml) induced similar effects as did MSC-CM. On the basis of a previous study, recombinant CX3CL1 (rCX3CL1, 10 ng/ml) was also tested. The rTGF-β and rCX3CL1 were applied for 24 hours. rTGF-β rescued the reduced CX3CR1 expression (A) and inhibited the increased gene expression of TNF-α (B), IL-1β (C), IL-6 (D), and iNOS (E), similar to MSC-CM in LPS-stimulated microglia. rCX3CL1 (10 ng/ml) also inhibited IL-1β and iNOS expression and restored CX3CR1 expression but did not affect TNF-α and IL-6 expression. The data are means ± SEM of three independent experiments. ∗, p < .05; ∗∗∗, p < .001; in comparison with control. §, p < .05; §§§, p < .001; in comparison with LPS. ††, p < .05; †††, p < .001; compared between two groups. Abbreviations: iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; RQ, relative quantity; TGF, transforming growth factor; TNF, tumor necrosis factor.

    Techniques Used: Recombinant, Expressing, Gene Expression, Comparison, Control

    MSC-CM inhibits the NF-κB pathway and rescues CX3CR1 expression via the TGF-β signaling pathway in LPS-stimulated microglia. (A): LPS increased p-IκB expression and reduced CX3CR1 expression and Smad2/3 phosphorylation by Western blot. MSC-CM restored these changes in LPS-stimulated microglia. However, the effect of MSC-CM was abolished when TGF-βR inhibitor was applied with MSC-CM. TGF-βR inhibitor alone did not affect CX3CR1 expression, IκB, or Smad2/3 phosphorylation in primary cultured microglia. The p-IκB (B), p-smad2 (C), p-smad3 (D), and CX3CR1 (E) expression levels were measured using densitometry. The data are means ± SEM of three independent experiments. ∗, p < .05, compared with control. §, p < .05; §§, p < .01; §§§, p < .001; in comparison with LPS. †, p < .05; ††, p < .001; compared between two groups. Abbreviations: CON, control; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; TGF, transforming growth factor.
    Figure Legend Snippet: MSC-CM inhibits the NF-κB pathway and rescues CX3CR1 expression via the TGF-β signaling pathway in LPS-stimulated microglia. (A): LPS increased p-IκB expression and reduced CX3CR1 expression and Smad2/3 phosphorylation by Western blot. MSC-CM restored these changes in LPS-stimulated microglia. However, the effect of MSC-CM was abolished when TGF-βR inhibitor was applied with MSC-CM. TGF-βR inhibitor alone did not affect CX3CR1 expression, IκB, or Smad2/3 phosphorylation in primary cultured microglia. The p-IκB (B), p-smad2 (C), p-smad3 (D), and CX3CR1 (E) expression levels were measured using densitometry. The data are means ± SEM of three independent experiments. ∗, p < .05, compared with control. §, p < .05; §§, p < .01; §§§, p < .001; in comparison with LPS. †, p < .05; ††, p < .001; compared between two groups. Abbreviations: CON, control; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; TGF, transforming growth factor.

    Techniques Used: Expressing, Phospho-proteomics, Western Blot, Cell Culture, Control, Comparison



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    rat msc conditioned media  (Thermo Fisher)
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    Thermo Fisher rat msc conditioned media
    <t>MSC-CM</t> suppressed the inflammatory reaction and enhanced phagocytosis in LPS-stimulated microglia. (A): To determine the effect of MSC-CM on the level of cytokines in LPS-stimulated microglia, we applied MSC-CM for 24, 48, and 72 hours to LPS-stimulated microglia. LPS increased mRNA expression of TNF-α, IL-1β, IL-6, and iNOS, and nitrate secretion in LPS-stimulated microglia, whereas MSC-CM inhibited the inflammatory reaction. (B): Immunostaining shows that MSC-CM significantly suppressed iNOS expression in LPS-stimulated microglia. The microglia and nuclei were stained with Iba-1 and DAPI, respectively. (C): LPS-stimulated microglia ingested more latex beads than did untreated controls, and the phagocytic activity of microglia was significantly enhanced when the MSC-CM was applied to LPS-stimulated microglia following LPS treatment. The data are means ± SEM of three independent experiments. ∗, p < .05; ∗∗, p < .01; ∗∗∗, p < .001; in comparison with control. §, p < .05; §§, p < .05; §§§, p < .001; in comparison with LPS. Abbreviations: CON, control; DAPI, 4′,6-diamidino-2-phenylindole; hr, hour; iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal <t>cell-conditioned</t> media; RQ, relative quantity; TNF, tumor necrosis factor.
    Rat Msc Conditioned Media, supplied by Thermo Fisher, 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/product/rat+msc+conditioned+media/pmc05070497-81-0-18?v=Thermo+Fisher
    Average 90 stars, based on 1 article reviews
    rat msc conditioned media - by Bioz Stars, 2026-08
    90/100 stars
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    MSC-CM suppressed the inflammatory reaction and enhanced phagocytosis in LPS-stimulated microglia. (A): To determine the effect of MSC-CM on the level of cytokines in LPS-stimulated microglia, we applied MSC-CM for 24, 48, and 72 hours to LPS-stimulated microglia. LPS increased mRNA expression of TNF-α, IL-1β, IL-6, and iNOS, and nitrate secretion in LPS-stimulated microglia, whereas MSC-CM inhibited the inflammatory reaction. (B): Immunostaining shows that MSC-CM significantly suppressed iNOS expression in LPS-stimulated microglia. The microglia and nuclei were stained with Iba-1 and DAPI, respectively. (C): LPS-stimulated microglia ingested more latex beads than did untreated controls, and the phagocytic activity of microglia was significantly enhanced when the MSC-CM was applied to LPS-stimulated microglia following LPS treatment. The data are means ± SEM of three independent experiments. ∗, p < .05; ∗∗, p < .01; ∗∗∗, p < .001; in comparison with control. §, p < .05; §§, p < .05; §§§, p < .001; in comparison with LPS. Abbreviations: CON, control; DAPI, 4′,6-diamidino-2-phenylindole; hr, hour; iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; RQ, relative quantity; TNF, tumor necrosis factor.

    Journal: Stem Cells Translational Medicine

    Article Title: Mesenchymal Stem Cells Modulate the Functional Properties of Microglia via TGF-β Secretion

    doi: 10.5966/sctm.2015-0217

    Figure Lengend Snippet: MSC-CM suppressed the inflammatory reaction and enhanced phagocytosis in LPS-stimulated microglia. (A): To determine the effect of MSC-CM on the level of cytokines in LPS-stimulated microglia, we applied MSC-CM for 24, 48, and 72 hours to LPS-stimulated microglia. LPS increased mRNA expression of TNF-α, IL-1β, IL-6, and iNOS, and nitrate secretion in LPS-stimulated microglia, whereas MSC-CM inhibited the inflammatory reaction. (B): Immunostaining shows that MSC-CM significantly suppressed iNOS expression in LPS-stimulated microglia. The microglia and nuclei were stained with Iba-1 and DAPI, respectively. (C): LPS-stimulated microglia ingested more latex beads than did untreated controls, and the phagocytic activity of microglia was significantly enhanced when the MSC-CM was applied to LPS-stimulated microglia following LPS treatment. The data are means ± SEM of three independent experiments. ∗, p < .05; ∗∗, p < .01; ∗∗∗, p < .001; in comparison with control. §, p < .05; §§, p < .05; §§§, p < .001; in comparison with LPS. Abbreviations: CON, control; DAPI, 4′,6-diamidino-2-phenylindole; hr, hour; iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; RQ, relative quantity; TNF, tumor necrosis factor.

    Article Snippet: Rat MSC Conditioned Media and Drug Treatment in Primary Cultured Microglia Rat (Sprague Dawley) MSCs were purchased from Gibco (#S1601-100; Thermo Fisher Scientific).

    Techniques: Expressing, Immunostaining, Staining, Activity Assay, Comparison, Control

    MSC-CM restored CX3CR1, CD206, and CD200R expression in LPS-stimulated microglia. To investigate the effect of MSC-CM on the expression of CD86, CX3CR1, CD206, and CD200R in LPS-stimulated microglia, we performed flow cytometry. (A): Microglia were treated with LPS for 24 hours and MSC-CM was incubated for 72 hours. The isotype control was indicated as gray color. Control group (red), LPS group (green), and LPS + MSC-CM (blue) were indicated, respectively. The table box beside the diagram indicates the percentage of positive cells in each group. (B): Rather than divide this microglia population further by imposing strict boundaries on a continuous expression pattern, we then determined MFI of the entire remaining population for each marker, and we calculated the M2:M1 MFI ratio (CX3CR1, CD206, or CD200R/CD86). MSC-CM restored CX3CR1, CD206, and CD200R MFI, whereas it inhibited the CD86 in LPS-stimulated microglia, showing the normalization of M2:M1 MFI ratio. (C): Immunofluorescence results showed a similar pattern with flow cytometry analysis. Microglia was stained with Iba-1 and DAPI, respectively. (D): MSC-CM restored mRNA expression of CX3CR1 and CD206 in LPS-stimulated microglia following to incubation time. The data are means ± SEM of three independent experiments. §, p < .001 in comparison with control (CON); §§, p < .01; §§§, p < .001 in comparison with LPS. Abbreviations: CON, control; DAPI, 4′,6-diamidino-2-phenylindole; hr, hour; LPS, lipopolysaccharide; MFI, mean fluorescence intensity; MSC-CM, mesenchymal stromal cell-conditioned media; RQ, relative quantity.

    Journal: Stem Cells Translational Medicine

    Article Title: Mesenchymal Stem Cells Modulate the Functional Properties of Microglia via TGF-β Secretion

    doi: 10.5966/sctm.2015-0217

    Figure Lengend Snippet: MSC-CM restored CX3CR1, CD206, and CD200R expression in LPS-stimulated microglia. To investigate the effect of MSC-CM on the expression of CD86, CX3CR1, CD206, and CD200R in LPS-stimulated microglia, we performed flow cytometry. (A): Microglia were treated with LPS for 24 hours and MSC-CM was incubated for 72 hours. The isotype control was indicated as gray color. Control group (red), LPS group (green), and LPS + MSC-CM (blue) were indicated, respectively. The table box beside the diagram indicates the percentage of positive cells in each group. (B): Rather than divide this microglia population further by imposing strict boundaries on a continuous expression pattern, we then determined MFI of the entire remaining population for each marker, and we calculated the M2:M1 MFI ratio (CX3CR1, CD206, or CD200R/CD86). MSC-CM restored CX3CR1, CD206, and CD200R MFI, whereas it inhibited the CD86 in LPS-stimulated microglia, showing the normalization of M2:M1 MFI ratio. (C): Immunofluorescence results showed a similar pattern with flow cytometry analysis. Microglia was stained with Iba-1 and DAPI, respectively. (D): MSC-CM restored mRNA expression of CX3CR1 and CD206 in LPS-stimulated microglia following to incubation time. The data are means ± SEM of three independent experiments. §, p < .001 in comparison with control (CON); §§, p < .01; §§§, p < .001 in comparison with LPS. Abbreviations: CON, control; DAPI, 4′,6-diamidino-2-phenylindole; hr, hour; LPS, lipopolysaccharide; MFI, mean fluorescence intensity; MSC-CM, mesenchymal stromal cell-conditioned media; RQ, relative quantity.

    Article Snippet: Rat MSC Conditioned Media and Drug Treatment in Primary Cultured Microglia Rat (Sprague Dawley) MSCs were purchased from Gibco (#S1601-100; Thermo Fisher Scientific).

    Techniques: Expressing, Flow Cytometry, Incubation, Control, Marker, Immunofluorescence, Staining, Comparison, Fluorescence

    TGF-β mediates the effect of MSC-CM on LPS-stimulated microglia. To determine which soluble factor was involved in the effect of MSC-CM on LPS-stimulated microglia, we treated LPS-stimulated microglia with MSC-CM + a TGF-βR inhibitor (A), a CX3CL1 antibody (B), or siMSC-CM (siMSC-CM indicates conditioned medium that was obtained from TGF-β siRNA-transfected MSCs) (C). LPS increased TNF-α, IL-1β, IL-6, and iNOS mRNA levels, and MSC-CM inhibited the increased cytokines and led to CX3CR1 restoration. TGF-β inhibition abolished the effect of MSC-CM on LPS-stimulated microglia, whereas CX3CL1 antibody did not. siMSC-CM did not affect LPS-stimulated microglia. The data are means ± SEM of three independent experiments. ∗, p < .01; ∗∗, p < .001; in comparison with control (CON). §, p < .05; §§, p < .01; §§§, p < .001; in comparison with LPS. †, p < .05; ††, p < .01; †††, p < .001; in comparison with two groups (LPS + MSC-CM vs. LPS + MSC-CM + antibody/inhibitor or siMSC-CM). Abbreviations: CON, control; iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; n.s., not significant; RQ, relative quantity; TGF, transforming growth factor; TNF, tumor necrosis factor.

    Journal: Stem Cells Translational Medicine

    Article Title: Mesenchymal Stem Cells Modulate the Functional Properties of Microglia via TGF-β Secretion

    doi: 10.5966/sctm.2015-0217

    Figure Lengend Snippet: TGF-β mediates the effect of MSC-CM on LPS-stimulated microglia. To determine which soluble factor was involved in the effect of MSC-CM on LPS-stimulated microglia, we treated LPS-stimulated microglia with MSC-CM + a TGF-βR inhibitor (A), a CX3CL1 antibody (B), or siMSC-CM (siMSC-CM indicates conditioned medium that was obtained from TGF-β siRNA-transfected MSCs) (C). LPS increased TNF-α, IL-1β, IL-6, and iNOS mRNA levels, and MSC-CM inhibited the increased cytokines and led to CX3CR1 restoration. TGF-β inhibition abolished the effect of MSC-CM on LPS-stimulated microglia, whereas CX3CL1 antibody did not. siMSC-CM did not affect LPS-stimulated microglia. The data are means ± SEM of three independent experiments. ∗, p < .01; ∗∗, p < .001; in comparison with control (CON). §, p < .05; §§, p < .01; §§§, p < .001; in comparison with LPS. †, p < .05; ††, p < .01; †††, p < .001; in comparison with two groups (LPS + MSC-CM vs. LPS + MSC-CM + antibody/inhibitor or siMSC-CM). Abbreviations: CON, control; iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; n.s., not significant; RQ, relative quantity; TGF, transforming growth factor; TNF, tumor necrosis factor.

    Article Snippet: Rat MSC Conditioned Media and Drug Treatment in Primary Cultured Microglia Rat (Sprague Dawley) MSCs were purchased from Gibco (#S1601-100; Thermo Fisher Scientific).

    Techniques: Transfection, Inhibition, Comparison, Control

    TGF-βR inhibition alone can abolish the effect of MSC-CM in LPS-stimulated microglia. To investigate whether CX3CL1 plays a role in the anti-inflammatory effect of TGF-β in MSC-CM, we added a CX3CL1 antibody, a TGF-βR inhibitor, and both together to MSC-CM. LPS reduced CX3CR1 (A) and increased TNF-α mRNA expression (B), IL-1β mRNA expression (C), IL-6 mRNA expression (D), and iNOS mRNA expression (E), and. MSC-CM rescued these changes. The TGF-βR inhibitor alone showed the same effect on LPS-stimulated microglia, regardless of the presence of the CX3CL1 antibody. When the TGF-βR inhibitor was added to the MSC-CM, the effect of the MSC-CM on TNF-α, IL-1β, IL-6, iNOS, and CX3CR1 was abolished. The data are means ± SEM of three independent experiments. ∗, p < .05; ∗∗, p < .01; ∗∗∗, p < .001; in comparison with control. §, p < .05; §§, p < .01; §§§, p < .001; in comparison with LPS. †, p < .05; †††, p < .001; compared between two groups. Abbreviations: iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; RQ, relative quantity; TGF, transforming growth factor; TNF, tumor necrosis factor.

    Journal: Stem Cells Translational Medicine

    Article Title: Mesenchymal Stem Cells Modulate the Functional Properties of Microglia via TGF-β Secretion

    doi: 10.5966/sctm.2015-0217

    Figure Lengend Snippet: TGF-βR inhibition alone can abolish the effect of MSC-CM in LPS-stimulated microglia. To investigate whether CX3CL1 plays a role in the anti-inflammatory effect of TGF-β in MSC-CM, we added a CX3CL1 antibody, a TGF-βR inhibitor, and both together to MSC-CM. LPS reduced CX3CR1 (A) and increased TNF-α mRNA expression (B), IL-1β mRNA expression (C), IL-6 mRNA expression (D), and iNOS mRNA expression (E), and. MSC-CM rescued these changes. The TGF-βR inhibitor alone showed the same effect on LPS-stimulated microglia, regardless of the presence of the CX3CL1 antibody. When the TGF-βR inhibitor was added to the MSC-CM, the effect of the MSC-CM on TNF-α, IL-1β, IL-6, iNOS, and CX3CR1 was abolished. The data are means ± SEM of three independent experiments. ∗, p < .05; ∗∗, p < .01; ∗∗∗, p < .001; in comparison with control. §, p < .05; §§, p < .01; §§§, p < .001; in comparison with LPS. †, p < .05; †††, p < .001; compared between two groups. Abbreviations: iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; RQ, relative quantity; TGF, transforming growth factor; TNF, tumor necrosis factor.

    Article Snippet: Rat MSC Conditioned Media and Drug Treatment in Primary Cultured Microglia Rat (Sprague Dawley) MSCs were purchased from Gibco (#S1601-100; Thermo Fisher Scientific).

    Techniques: Inhibition, Expressing, Comparison, Control

    Recombinant TGF-β showed an effect similar to that of MSC-CM on LPS-stimulated microglia. Recombinant TGF-β (rTGF-β, 10 ng/ml) induced similar effects as did MSC-CM. On the basis of a previous study, recombinant CX3CL1 (rCX3CL1, 10 ng/ml) was also tested. The rTGF-β and rCX3CL1 were applied for 24 hours. rTGF-β rescued the reduced CX3CR1 expression (A) and inhibited the increased gene expression of TNF-α (B), IL-1β (C), IL-6 (D), and iNOS (E), similar to MSC-CM in LPS-stimulated microglia. rCX3CL1 (10 ng/ml) also inhibited IL-1β and iNOS expression and restored CX3CR1 expression but did not affect TNF-α and IL-6 expression. The data are means ± SEM of three independent experiments. ∗, p < .05; ∗∗∗, p < .001; in comparison with control. §, p < .05; §§§, p < .001; in comparison with LPS. ††, p < .05; †††, p < .001; compared between two groups. Abbreviations: iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; RQ, relative quantity; TGF, transforming growth factor; TNF, tumor necrosis factor.

    Journal: Stem Cells Translational Medicine

    Article Title: Mesenchymal Stem Cells Modulate the Functional Properties of Microglia via TGF-β Secretion

    doi: 10.5966/sctm.2015-0217

    Figure Lengend Snippet: Recombinant TGF-β showed an effect similar to that of MSC-CM on LPS-stimulated microglia. Recombinant TGF-β (rTGF-β, 10 ng/ml) induced similar effects as did MSC-CM. On the basis of a previous study, recombinant CX3CL1 (rCX3CL1, 10 ng/ml) was also tested. The rTGF-β and rCX3CL1 were applied for 24 hours. rTGF-β rescued the reduced CX3CR1 expression (A) and inhibited the increased gene expression of TNF-α (B), IL-1β (C), IL-6 (D), and iNOS (E), similar to MSC-CM in LPS-stimulated microglia. rCX3CL1 (10 ng/ml) also inhibited IL-1β and iNOS expression and restored CX3CR1 expression but did not affect TNF-α and IL-6 expression. The data are means ± SEM of three independent experiments. ∗, p < .05; ∗∗∗, p < .001; in comparison with control. §, p < .05; §§§, p < .001; in comparison with LPS. ††, p < .05; †††, p < .001; compared between two groups. Abbreviations: iNOS, inducible nitric oxide; IL, interleukin; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; RQ, relative quantity; TGF, transforming growth factor; TNF, tumor necrosis factor.

    Article Snippet: Rat MSC Conditioned Media and Drug Treatment in Primary Cultured Microglia Rat (Sprague Dawley) MSCs were purchased from Gibco (#S1601-100; Thermo Fisher Scientific).

    Techniques: Recombinant, Expressing, Gene Expression, Comparison, Control

    MSC-CM inhibits the NF-κB pathway and rescues CX3CR1 expression via the TGF-β signaling pathway in LPS-stimulated microglia. (A): LPS increased p-IκB expression and reduced CX3CR1 expression and Smad2/3 phosphorylation by Western blot. MSC-CM restored these changes in LPS-stimulated microglia. However, the effect of MSC-CM was abolished when TGF-βR inhibitor was applied with MSC-CM. TGF-βR inhibitor alone did not affect CX3CR1 expression, IκB, or Smad2/3 phosphorylation in primary cultured microglia. The p-IκB (B), p-smad2 (C), p-smad3 (D), and CX3CR1 (E) expression levels were measured using densitometry. The data are means ± SEM of three independent experiments. ∗, p < .05, compared with control. §, p < .05; §§, p < .01; §§§, p < .001; in comparison with LPS. †, p < .05; ††, p < .001; compared between two groups. Abbreviations: CON, control; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; TGF, transforming growth factor.

    Journal: Stem Cells Translational Medicine

    Article Title: Mesenchymal Stem Cells Modulate the Functional Properties of Microglia via TGF-β Secretion

    doi: 10.5966/sctm.2015-0217

    Figure Lengend Snippet: MSC-CM inhibits the NF-κB pathway and rescues CX3CR1 expression via the TGF-β signaling pathway in LPS-stimulated microglia. (A): LPS increased p-IκB expression and reduced CX3CR1 expression and Smad2/3 phosphorylation by Western blot. MSC-CM restored these changes in LPS-stimulated microglia. However, the effect of MSC-CM was abolished when TGF-βR inhibitor was applied with MSC-CM. TGF-βR inhibitor alone did not affect CX3CR1 expression, IκB, or Smad2/3 phosphorylation in primary cultured microglia. The p-IκB (B), p-smad2 (C), p-smad3 (D), and CX3CR1 (E) expression levels were measured using densitometry. The data are means ± SEM of three independent experiments. ∗, p < .05, compared with control. §, p < .05; §§, p < .01; §§§, p < .001; in comparison with LPS. †, p < .05; ††, p < .001; compared between two groups. Abbreviations: CON, control; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; LPS, lipopolysaccharide; MSC-CM, mesenchymal stromal cell-conditioned media; TGF, transforming growth factor.

    Article Snippet: Rat MSC Conditioned Media and Drug Treatment in Primary Cultured Microglia Rat (Sprague Dawley) MSCs were purchased from Gibco (#S1601-100; Thermo Fisher Scientific).

    Techniques: Expressing, Phospho-proteomics, Western Blot, Cell Culture, Control, Comparison