Figure 6.

Generation of a vascularized 3D cardiac construct by adult stem cells grown in dynamic conditions onto gelatin porous scaffolds. Schematic illustration of the protocol used to produce vascularized 3D cardiac construct (A). Gelatin porous sponges were dipped in diluted Matrigel™ and then switched to endothelial differentiation medium (EDM). Therefore, hMSCs were allowed to colonize the scaffold and differentiate toward the endothelial phenotype for 4 days. Cardiac TNT-GFP progenitor cells were pre-committed for 2 weeks with cardiac differentiation medium (CDM) on TCPS, and then cultured on vascularized scaffold for further 7 days in CDM and in a perfusive modular bioreactor. Human cells colonize the scaffold inner layers as shown by nuclei staining at different depths (B). Side view of cellularized scaffolds cultured under static or dynamic conditions (C); infiltration of GFP- (cardiomyocyte-like cells, green) and VCAM-1-positive cells (endothelial-like cells, red) into scaffold is improved by dynamic culture. Immunohistochemistry analysis of the colonized scaffolds shows the massive infiltration of VCAM-1, CD144 cells in the core of the construct. Higher magnification images (D) shows the VCAM-1-positive cells aligned in tube-like structures around the pores and contacting GFP-positive cells (E,F). S, scaffold.