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Co-transplantation of PEP with SC-β cells delays NK-cell recruitment and xenograft rejection. ( A ) Human insulin levels in plasma from C57BL/6 mice measured by human-specific insulin <t>ELISA</t> at days 2, 5, 14, and 21 post-transplantations. ( B ) Quantification of insulin + cells within the left kidney capsule at various time points following SC-β cell transplantation. ( C ) Immune cell infiltration in xenogeneic grafts was quantified based on marker expression for CD45 + leukocytes, Ly6G + neutrophils, F4/80 + macrophages, CD11b + myeloid cells, CD11c + dendritic cells, B220 + B cells, CD3 + T cells, and NK1.1 + natural killer cells. ( D ) Representative immunofluorescence images showing insulin (green) and NK1.1 (red) staining, with nuclei counterstained with DAPI (blue). Images were acquired at 20× magnification; scale bars represent 50 μm. Data are presented as mean ± SEM. Statistical analysis was performed using unpaired Student’s t -test for two-group comparisons and two-way ANOVA followed by multiple-comparison testing for multi-group comparisons. Statistical significance is indicated as * p < 0.05, **** p < 0.0001.
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Co-transplantation of PEP with SC-β cells delays NK-cell recruitment and xenograft rejection. ( A ) Human insulin levels in plasma from C57BL/6 mice measured by human-specific insulin <t>ELISA</t> at days 2, 5, 14, and 21 post-transplantations. ( B ) Quantification of insulin + cells within the left kidney capsule at various time points following SC-β cell transplantation. ( C ) Immune cell infiltration in xenogeneic grafts was quantified based on marker expression for CD45 + leukocytes, Ly6G + neutrophils, F4/80 + macrophages, CD11b + myeloid cells, CD11c + dendritic cells, B220 + B cells, CD3 + T cells, and NK1.1 + natural killer cells. ( D ) Representative immunofluorescence images showing insulin (green) and NK1.1 (red) staining, with nuclei counterstained with DAPI (blue). Images were acquired at 20× magnification; scale bars represent 50 μm. Data are presented as mean ± SEM. Statistical analysis was performed using unpaired Student’s t -test for two-group comparisons and two-way ANOVA followed by multiple-comparison testing for multi-group comparisons. Statistical significance is indicated as * p < 0.05, **** p < 0.0001.
Ultrasensitive Insulin Elisa Kit, supplied by ALPCO, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Co-transplantation of PEP with SC-β cells delays NK-cell recruitment and xenograft rejection. ( A ) Human insulin levels in plasma from C57BL/6 mice measured by human-specific insulin ELISA at days 2, 5, 14, and 21 post-transplantations. ( B ) Quantification of insulin + cells within the left kidney capsule at various time points following SC-β cell transplantation. ( C ) Immune cell infiltration in xenogeneic grafts was quantified based on marker expression for CD45 + leukocytes, Ly6G + neutrophils, F4/80 + macrophages, CD11b + myeloid cells, CD11c + dendritic cells, B220 + B cells, CD3 + T cells, and NK1.1 + natural killer cells. ( D ) Representative immunofluorescence images showing insulin (green) and NK1.1 (red) staining, with nuclei counterstained with DAPI (blue). Images were acquired at 20× magnification; scale bars represent 50 μm. Data are presented as mean ± SEM. Statistical analysis was performed using unpaired Student’s t -test for two-group comparisons and two-way ANOVA followed by multiple-comparison testing for multi-group comparisons. Statistical significance is indicated as * p < 0.05, **** p < 0.0001.

Journal: Cells

Article Title: Platelet-Derived Exosome Product Prolongs Stem Cell-Derived β-Cell Graft Survival and Is Associated with Reduced Nk Cell Infiltration and Immunomodulation

doi: 10.3390/cells15151367

Figure Lengend Snippet: Co-transplantation of PEP with SC-β cells delays NK-cell recruitment and xenograft rejection. ( A ) Human insulin levels in plasma from C57BL/6 mice measured by human-specific insulin ELISA at days 2, 5, 14, and 21 post-transplantations. ( B ) Quantification of insulin + cells within the left kidney capsule at various time points following SC-β cell transplantation. ( C ) Immune cell infiltration in xenogeneic grafts was quantified based on marker expression for CD45 + leukocytes, Ly6G + neutrophils, F4/80 + macrophages, CD11b + myeloid cells, CD11c + dendritic cells, B220 + B cells, CD3 + T cells, and NK1.1 + natural killer cells. ( D ) Representative immunofluorescence images showing insulin (green) and NK1.1 (red) staining, with nuclei counterstained with DAPI (blue). Images were acquired at 20× magnification; scale bars represent 50 μm. Data are presented as mean ± SEM. Statistical analysis was performed using unpaired Student’s t -test for two-group comparisons and two-way ANOVA followed by multiple-comparison testing for multi-group comparisons. Statistical significance is indicated as * p < 0.05, **** p < 0.0001.

Article Snippet: Plasma was isolated from whole blood and used to quantify human insulin levels using a Human Ultrasensitive Insulin ELISA kit (ALPCO, Salem, NH, USA; Cat. No. 80-INSHUU-E10).

Techniques: Transplantation Assay, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Marker, Expressing, Immunofluorescence, Staining, Comparison

PEP delays xenograft rejection and limits NK-cell recruitment across multiple administration routes. ( A ) Schematic illustration of SC-β cell transplantation with different PEP administration strategies. Group 1: SC-β cells only; Group 2: SC-β cells pretreated in vitro with 10% PEP for 24 h before transplantation; Group 3: SC-β cells co-transplanted with 100% PEP (15 µL); Group 4: systemic PEP delivery via intraperitoneal (IP) injection for 3 days (25% PEP, 200 µL); Group 5: saline control. ( B ) Human insulin levels in plasma from C57BL/6 mice at the indicated time points following transplantation, measured by human-specific insulin ELISA. Data are presented as mean ± SEM. Statistical comparisons were performed among treatment groups at each time point; no statistically significant differences were detected. ( C ) Heatmap of relative mRNA expression of selected immune-related genes in graft-bearing kidneys harvested on day 14. Genes are organized by functional category, including general immune markers, T-cell-associated markers, macrophage-associated markers, chemokines/cytokines, inflammasome-related genes, and complement-associated genes. Expression values are shown as log 2 (2 −ΔΔCT ) relative to sham controls.

Journal: Cells

Article Title: Platelet-Derived Exosome Product Prolongs Stem Cell-Derived β-Cell Graft Survival and Is Associated with Reduced Nk Cell Infiltration and Immunomodulation

doi: 10.3390/cells15151367

Figure Lengend Snippet: PEP delays xenograft rejection and limits NK-cell recruitment across multiple administration routes. ( A ) Schematic illustration of SC-β cell transplantation with different PEP administration strategies. Group 1: SC-β cells only; Group 2: SC-β cells pretreated in vitro with 10% PEP for 24 h before transplantation; Group 3: SC-β cells co-transplanted with 100% PEP (15 µL); Group 4: systemic PEP delivery via intraperitoneal (IP) injection for 3 days (25% PEP, 200 µL); Group 5: saline control. ( B ) Human insulin levels in plasma from C57BL/6 mice at the indicated time points following transplantation, measured by human-specific insulin ELISA. Data are presented as mean ± SEM. Statistical comparisons were performed among treatment groups at each time point; no statistically significant differences were detected. ( C ) Heatmap of relative mRNA expression of selected immune-related genes in graft-bearing kidneys harvested on day 14. Genes are organized by functional category, including general immune markers, T-cell-associated markers, macrophage-associated markers, chemokines/cytokines, inflammasome-related genes, and complement-associated genes. Expression values are shown as log 2 (2 −ΔΔCT ) relative to sham controls.

Article Snippet: Plasma was isolated from whole blood and used to quantify human insulin levels using a Human Ultrasensitive Insulin ELISA kit (ALPCO, Salem, NH, USA; Cat. No. 80-INSHUU-E10).

Techniques: Transplantation Assay, In Vitro, Injection, Saline, Control, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Expressing, Functional Assay