title: Adipose tissue-derived stem cells display a proangiogenic phenotype on 3D scaffolds.
abstract: Ischemic heart disease is the leading cause of death worldwide. Recent studies suggest that adipose tissue-derived stem cells (ASCs) can be used as a potential source for cardiovascular tissue engineering due to their ability to differentiate along the cardiovascular lineage and to adopt a proangiogenic phenotype. To understand better ASCs' biology, we used a novel 3D culture device. ASCs' and b.END-3 endothelial cell proliferation, migration, and vessel morphogenesis were significantly enhanced compared to 2D culturing techniques. ASCs were isolated from inguinal fat pads of 6-week-old GFP+/BLI+ mice. Early passage ASCs cells (P3-P4), PKH26-labeled murine b.END-3 cells or a co-culture of ASCs and b.END-3 cells were seeded at a density of 1 × 10(5) on three different surface configurations: (a) a 2D surface of tissue culture plastic, (b) Matrigel, and (c) a highly porous 3D scaffold fabricated from inert polystyrene. VEGF expression, cell proliferation, and tubulization, were assessed using optical microscopy, fluorescence microscopy, 3D confocal microscopy, and SEM imaging (n = 6). Increased VEGF levels were seen in conditioned media harvested from co-cultures of ASCs and b.END-3 on either Matrigel or a 3D matrix. Fluorescence, confocal, SEM, bioluminescence revealed improved cell, proliferation, and tubule formation for cells seeded on the 3D polystyrene matrix. Collectively, these data demonstrate that co-culturing ASCs with endothelial cells in a 3D matrix environment enable us to generate prevascularized tissue-engineered constructs. This can potentially help us to surpass the tissue thickness limitations faced by the tissue engineering community today.
authors.affiliation: Department of Cardiothoracic Surgery, Stanford University School of Medicine, California, USA.
authors.forename: Evgenios A
authors.initials: EA
authors.lastname: Neofytou
authors.forename: Edwin
authors.initials: E
authors.lastname: Chang
authors.forename: Bhagat
authors.initials: B
authors.lastname: Patlola
authors.forename: Lydia-Marie
authors.initials: LM
authors.lastname: Joubert
authors.forename: Jayakumar
authors.initials: J
authors.lastname: Rajadas
authors.forename: Sanjiv S
authors.initials: SS
authors.lastname: Gambhir
authors.forename: Zhen
authors.initials: Z
authors.lastname: Cheng
authors.forename: Robert C
authors.initials: RC
authors.lastname: Robbins
authors.forename: Ramin E
authors.initials: RE
authors.lastname: Beygui
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D042783:Endothelial Cells / Q000166:cytology; D051379:Mice; D018919:Neovascularization, Physiologic; D016062:Porosity; D013234:Stem Cells / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*