Journal: Scientific Reports
Article Title: Surface modification of pig endothelial cells with a branched heparin conjugate improves their compatibility with human blood
doi: 10.1038/s41598-017-04898-w
Figure Lengend Snippet: ( A–C ) CHC coating of PAEC reduces complement deposition, cellular activation and coagulation induced by human plasma. Uncoated and CHC-coated PAEC were treated for 4 hrs with 10% human plasma, and assessed by ELISA for ( A ) the deposition of activated complement C5b-9 and ( B ) the expression of the adhesion molecule E-selectin ( B ). ( C ) Supernatants from this assay were measured for D-dimers concentration. CHC coating significantly protected WT PAEC. Uncoated GTKO.hCD46.hTBM PAEC were significantly protected compared to uncoated WT PAEC; coating with CHC further reduced complement deposition and cellular activation, although this was not statistically significant. ( D,F ) CHC coating of hTNFα-activated PAEC reduces complement deposition, cellular activation and coagulation induced by human plasma. WT and GTKO.hCD46.hTBM PAEC were treated with hTNFα (100 ng/ml) for 2 hrs to induce cellular activation and shedding of the endothelial glycocalyx. After, PAEC were coated with CHC and incubated for 4 hrs with 10% human plasma, and assessed for ( D ) complement C5b-9 deposition and ( E ) endothelial E-selectin expression as well as ( F ) D-dimer levels in the supernatants. CHC coating significantly protected hTNFα-activated WT and GTKO.hCD46.hTBM PAEC compared to uncoated PAEC. Significance was measured by one-way ANOVA with Bonferroni correction (*p < 0.05, **p < 0.01, ***p < 0.001). Data are mean ± SD of three independent experiments.
Article Snippet: Rabbit anti-human C3b/c FITC (A0062; Dako), mouse anti-human C5b-9 FITC (clone aE11; HM2167F, Hycult Biotech), and mouse anti-human CD62E FITC (clone 1.2B6, MCA883F, AbD Serotec) were diluted in PBS/1% BSA and incubated for 60 min at RT followed by three washes.
Techniques: Activation Assay, Coagulation, Enzyme-linked Immunosorbent Assay, Expressing, Concentration Assay, Incubation
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