Figure 2
From: Evaluation of a Miniaturized Biologically Vascularized Scaffold in vitro and in vivo
Re-endothelialization of the acellular mBioVaSc’s vascular tree. The preserved vascular system of the acellular mBioVaSc was repopulated with mvECs and cultured in a bioreactor mimicking physiological perfusion. The repopulation of the vessels was determined by live imaging, immunohistochemistry, metabolic activity, viability and functionality displaying branching capillaries in the luminal wall lined with endothelial cells. (a) Live imaging of a mBioVaSc segment with ECs expressing GFP and RFP infused through the arterial and venous pedicle, respectively. (b) Metabolic activity of EC-seeded mBioVaSc by colorimetric MTT assay. (c) Life-dead staining differentiating viable (FDA) and apoptotic/necrotic (PI) endothelial cells. (d) Light sheet microscopical visualization of capillaries inside opposing luminal walls. (e) Immunohistological detection of the endothelial marker CD31 in a cross section of the luminal revascularized capillary structures (highlighted by arrowheads). (f) Whole mount immunofluorescence staining of the EC-marker CD31 on the luminal surface. (g) AcLDL uptake by ECs in the capillary bed of the mBioVaSc. (h) IHC staining of vWF in a cross section (highlighted by arrowheads). (i) Immunofluorescence (IF) whole mount staining of vWF. Scale bars are 3 mm for (a); 1 mm for (b,c,f,i); 200 µm for (d,e,h); 100 µm for (g).
