a hematoxylin and eosin staining of the patient surgical specimen showing high-grade polymorphic UPS cells (light blue stroma) infiltrating necrotic tissue, 10 x and 20 x magnification. b hematoxylin and eosin staining of the cytospunned tumor cells from the patient-derived UPS primary culture S1, 10 x and 20 x magnification. c hematoxylin and eosin staining of the cytospunned tumor cells (light blue spots) from the patient-derived UPS primary culture S1 cultured within 3D collagen-based scaffold system, 10 x and 20 x magnification. d Heat map comparisons of the relative gene expression of selected tumor-associated markers tgf-b, slug, snail, mmp9 between UPS patient surgical specimen, S1 patient-derived 2D standard monolayer primary culture and S1 patient-derived 3D primary culture system. e hematoxylin and eosin staining of the patient surgical specimen showing low and high grade UPS and L-sarcoma (light blue stroma) infiltrating adipose tissue, 10x and 20× magnification. f hematoxylin and eosin staining of the cytospunned tumor cells of patient-derived primary cultures UPS and L-sarcoma case series, 10 x and 20 x magnification

a Chemobiogram analysis of S1 primary culture seeded in 2D and 3D-collagen based scaffold and exposed to chemotherapeutics agent, untreated cells were used as control. b Representative images of 2D and 3D-collagen based scaffold S1 primary culture exposed to the tested drugs. c DNA fragmentation analysis expressed as apoptotic cells % of 2D and 3D-collagen based scaffold S1 primary culture exposed to the tested drugs. d Representative images (dot plot) of DNA fragmentation analysis obtained thought flow cytometry

a Proteomic analysis of apoptotic- and anti-apoptotic-related proteins in 2D S1 primary culture exposed to the tested drugs. b Densitometric analysis of protein bands % normalized on the housekeeping vinculin. c Proteomic analysis of apoptotic- and anti-apoptotic-related proteins in 3D-collagen based scaffold S1 primary culture exposed to the tested drugs. d Densitometric analysis of protein bands % normalized on the housekeeping vinculin

a-c Relative expression of ECM-associated genes in 2D and 3D-collagen based scaffold S1 primary culture. d timp1/mmp2 relative expression ratio genes in 2D and 3D-collagen based scaffold S1 primary culture. e timp1/mmp9 relative expression ratio genes in 2D and 3D-collagen based scaffold S1 primary culture

a Representative fluorescence microscopy images of zebrafish embryos xenotrasnplanted with S1 cultured in standard monolayer culture (2D) and within 3D collagen-based scaffold (3D). Images of embryos untreated at 2 and 72 h post injection and exposed to trabectedin at 72 h post injection, scale bar 1000 μm. b Mean fluorescence signal of 2D and 3D xenotransplanted embryos, arbitrary units. c Tumor growth inhibition rate between 2D and 3D groups

Pharmacological profile of 2D and 3D-collagen based scaffold UPS and L-sarcoma primary culture case series. Primary cells were exposed to selected first- and second- line treatments (EPI, TRABE, ERI, DACA) for STS. Images of the surgical specimens used for the establishment of primary cultures are reported (a) S2 DDLPS. b S3 DDLPS. c S4 ALT/WDLPS. d S5 LMS. e S6 ALT/WDLPS. f S7 ALT/WDLPS. g S8 UPS. h DDLPS. i PLS