Journal: International Journal of Molecular Sciences

Article Title: Proinflammatory Cytokine Preconditioning Enhances the Therapeutic Potency of Different Types of MSCs in Inflammation

doi: 10.3390/ijms27094090

Figure Lengend Snippet: Cytomix increased the expression of immunomodulatory markers on BM-MSCs, iMSCs WT and iMSCs B2M KO. The gating strategy is shown in . ( A ) Histogram charts of the expression of the immunomodulatory markers on BM-MSCs, iMSCs WT and iMSCs B2M KO. ( B – H ) Cytomix significantly increased the expression levels of HLA-ABC on BM-MSCs and iMSCs WT, HLA-DR on BM-MSCs, CD54 on all types of MSCs, CD200 on iMSCs WT, CD119 and CD120b on BM-MSCs, and CD274 on iMSCs WT and iMSCs B2M KO. Representative results of three independent experiments are shown. Results are shown as mean ± SD ( n = 3 in each group) and analysed by two-way ANOVA with Šídák’s multiple comparisons test or Tukey’s multiple comparisons test. */**/***/**** represent comparisons of naïve vs. Cytomix in each type of MSC; $/$$$/$$$$ represent comparisons of BM-MSCs vs. iMSCs WT vs. iMSCs B2M KO in the naïve group; ##/###/#### represent comparisons of BM-MSCs vs. iMSCs WT vs. iMSCs B2M KO in the Cytomix group with significance levels of p < 0.05/0.01/0.001/0.0001, respectively. Nd represents not detectable.

Article Snippet: The cells were centrifuged at 300× g for 5 min and resuspended in 100 μL FACS buffer containing anti-human HLA-ABC-VioGreen, HLA-DR-VioBlue, CD54-APC, CD274-PE-Vio ® 615, CD119-FITC (Cat# 130-120-436, 130-111-794, 130-121-342, 130-122-811, 130-099-931; Miltenyi Biotec, Bergisch Gladbach, Germany), CD120b-PE and CD200-PE-CY7 antibodies (Cat# 358403, 399805; BioLegend, San Diego, CA, USA).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: GJA5 and ATP1A1 perturbations recapitulate inflammation-related beat irregularities in iPSC-based atrial myocardium tissue model

doi: 10.3389/fimmu.2025.1719392

Figure Lengend Snippet: Fabrication of 3D condensed tissues incorporating M1 macrophages. (A) Schematic of 3D tissue formation protocol, consisting of aCM, cfb, and monocyte preculture for 3 days (d-10), tissue seeding in 96 well plates (d-7) subsequent culture and M1 maturation for 7 days (d-7 to d0) and activation of M1 (d0). (B) Brightfield image of the injection molded pillar device used for seeding tissues. (C) Annotated representative brightfield image of condensed 3D tissue consisting of aCM+cfb. 9 D) Sharp electrode recordings of single cell aCMs in 3D tissues consisting of aCM+cfb (n=17/N=4) compared to 2D monolayer of aCM for (n=31/N=3), demonstrating significant increase in upstroke velocity for 3D tissues (Mann-Whitney test). (E) IF staining for cardiac marker cTnT and cardiac tissue resident macrophage marker CX3CR1 in aCM+cfb+M1 tissue. Zoomed-in image showing area marked by white rectangle, white arrows indicate macrophages. ****P<0.0001.

Article Snippet: Fixed aCM in suspension were co-stained with MLC2a REAfinityTM conjugated with APC (1:10, Miltenyi Biotec) and cTnT REAfinityTM conjugated with FITC (1:10, Miltenyi Biotec) antibodies and incubated for 15 min at RT.

Techniques: Activation Assay, Injection, Single Cell, MANN-WHITNEY, Staining, Marker

Journal: Frontiers in Immunology

Article Title: GJA5 and ATP1A1 perturbations recapitulate inflammation-related beat irregularities in iPSC-based atrial myocardium tissue model

doi: 10.3389/fimmu.2025.1719392

Figure Lengend Snippet: Fabrication of 3D condensed tissues incorporating M1 macrophages. (A) Schematic of 3D tissue formation protocol, consisting of aCM, cfb, and monocyte preculture for 3 days (d-10), tissue seeding in 96 well plates (d-7) subsequent culture and M1 maturation for 7 days (d-7 to d0) and activation of M1 (d0). (B) Brightfield image of the injection molded pillar device used for seeding tissues. (C) Annotated representative brightfield image of condensed 3D tissue consisting of aCM+cfb. 9 D) Sharp electrode recordings of single cell aCMs in 3D tissues consisting of aCM+cfb (n=17/N=4) compared to 2D monolayer of aCM for (n=31/N=3), demonstrating significant increase in upstroke velocity for 3D tissues (Mann-Whitney test). (E) IF staining for cardiac marker cTnT and cardiac tissue resident macrophage marker CX3CR1 in aCM+cfb+M1 tissue. Zoomed-in image showing area marked by white rectangle, white arrows indicate macrophages. ****P<0.0001.

Article Snippet: Tissues were incubated with primary antibodies (cTnT REAfinity conjugated FITC (1:100, Miltenyi Biotec)) and CX3CR1 rabbit anti-human (1H14L7) (1:250, Invitrogen) or Collagen I Polyclonal Antibody (1:100, ThermoFisher Scientific) overnight at 4 °C on a plate shaker.

Techniques: Activation Assay, Injection, Single Cell, MANN-WHITNEY, Staining, Marker