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. 2015 Jun 23;4(8):956–966. doi: 10.5966/sctm.2014-0259

Figure 4.

Figure 4.

Detection of implanted UCBMSCs and functional vascular structures induced in the infarcted myocardium and fibrin-cell patch interface. (A–C): Representative fluorescence confocal microscope images of a 4-week-old fibrin-cell patch showing, from left to right, red fluorescent human UCBMSCs expressing cytomegalovirus promoter-regulated monomeric red fluorescent protein (mRFP1) (A), green fluorescent UCBMSCs expressing CD31 promoter-regulated eGFP (B), and cells expressing CD31 protein (C). (D): Merged image of A, B, and C showing nuclei counterstained with Hoechst 33342. (E): Histogram showing the vessel density expressed as the percentage of the area occupied by GSLI B4+ cells in sham-UCBMSC, control-MI, MI-fibrin, and MI-UCBMSC mice. (F): GSLI B4 staining (white) showing vessel-like structures within the fibrin-cell patch at 4 weeks after infarction. (G): Vascular structures at the interface (dashed line) between the subjacent infarcted myocardium wound and engrafted construct. Arrowheads indicate a representative functional microvessel filled with fluorescein isothiocyanate-dextran (green) traversing the interface between the host myocardium and cell-seeded fibrin patch (top left). Red fluorescent UCBMSCs expressing mRFP1 frequently populated the fibrin patch but not the subjacent myocardium (top right). Cardiomyocytes, labeled using a specific anti-cardiac troponin I antibody (white), exclusively populated the myocardial side of the interface (bottom left). Superimposition of the three images showed nuclei counterstained with Hoechst 33342 and yellow color stain, resulting from the superposition of red fluorescent UBCMSCs and green fluorescent dextran (bottom right). Scale bars = 20 μm. Abbreviations: control-MI, myocardial infarction induced but no treatment; eGFP, enhanced green fluorescent protein; MI-fibrin, myocardial infarction treated with fibrin alone; MI-UCBMSC, myocardial infarction treated with umbilical cord blood mesenchymal stem cells; Sham-UCBMSC, no myocardial infarction but implantation of fibrin-cell patch.