Fig. 3

PDX model establishment from ccRCC injection. a Hematoxylin and Eosin staining of human tumor engrafted in murine models. A representative image of kidney PDX was reported. Microscope Nikon Eclipse E1000 10X and 20X (b) Graph reporting the percentage of patients who engrafted when orthotopically injected in mice and distributed following grading (3 G1; 7 G2; 13 G3; 7 G4). Eight mice for each patient were injected and all 18 tumors on 30 which were evaluated as engrafted developed tumor masses on ≥ of 6 mice. Tumors declared unable to engraft did not produce, at all, tumor masses. c Representative images (1 × 0.63) of the PDXs excised 90 days after injection by Stereomicroscope (Olympus SZX10,XCX50). One representative image for G2, G3 and G4 types was reported. d Representative images (1 × 0.63 and 1.25) of aberrant neo-angiogenesis formation in PDXs by Stereomicroscope (Olympus SZX10, XCX50 camera). e Hematoxylin and Eosin staining of PDXs versus parental primary patients. One representative tumor for each grade (G2, G3, G4) was reported. The staining was executed on OCT frozen samples. f Hematoxylin and Eosin and anti-PAX8, CD10, Vimentin and EMA staining were reported on formalin-fixed and paraffin- embedded parental primary, metastatic tissues and PDXs. Primary, metastatic and PDX tissues were obtained and shown from one representative patient. Microscope Nikon Eclipse 55i, magnification 20×. g Histogram showing the number of engrafting tumor populations evaluated over 30 injected patient samples and correlated with patient recurrence frequency calculated as development of metastases after surgery. Orange color represents recurrent (n = 7) and metastatic (n = 3) patients in the engrafted group (n = 18). Pink color represents recurrent patients (n = 2) in the non-engrafted group (n = 12) for a total of 30 injected samples