Fig. 6
a Immunohistochemical detection of GFP+ paMSC in heart sections. Heart sections were analyzed from two group III pigs (Heart 2 section) using Alexa 568-labeled anti-GFP (red) and DAPI (blue) counterstaining. Left, negative controls (DAPI alone) to establish tissue autofluorescence. Inset, enlarged image from one animal (dotted area). b IGF-1 expression in myocardial sections of a representative group III pig. Left, anti-IGF-1 staining developed with DAB immunohistochemistry in a representative section from a group III pig (paMSC-IGF- 1-GFP/HGF-Cherry). Right, negative control (no anti-IGF-1). c Graphical representation of pathological studies on inflammation, fibrosis, and new vessel formation in the collection of samples analyzed in all experimental groups studied. Values shown as the mean of various sections for each animal in each group (n = 4 animals/group; *p ≤0.05) (for criteria, see Methods). d Histological evaluation by H/E staining in group II pigs (paMSC-GFP+). Fibrosis is very marked in this group, with calcification and granulomatous areas. (D1, D2) Heart 2 section, animal 2; (D3) Heart 2 section, animal 1, in which we observed dystrophic calcifications surrounded by multinucleated giant cells. (D4) IVS section, animal 1 showed severe fibrosis accompanied by neovascularization. (D1) Thick arrow: area of calcification; thin arrow: lymphoplasmacytic granuloma; (D2) lymphoplasmacytic granuloma; (D3) thick arrow: area of calcification; thin arrow: giant cells; (D4) Arrows shows fibrosis. e Histological evaluation by H/E staining in group III pigs (paMSC-IGF-1/HGF). (E1, E2) Heart 2 section, animal 2 showed numerous granulomas with calcification in the fibrotic area. (E3) IVS section; animal 2 showed cartilaginous metaplasia. (E4) IVS section, animal 3, large areas of calcification and a severe fibrosis. (E1) Thick arrow: area of calcification; thin arrow: granuloma. Around fibrosis; (E2) arrows show granuloma; (E3) arrows indicate the cartilaginous metaplasia; (E4) thick arrow: area of calcification. The rest signals were assigned to fibrotic areas. GFP green fluorescent protein, HGF hepatocyte growth factor, IGF-1 insulin-like growth factor-1, IVS interventricular septum, LV left ventricular, paMSC pig mesenchymal stem cells from adipose tissue