Three-dimensional cell aggregates composed of HUVECs and cbMSCs for therapeutic neovascularization in a mouse model of hindlimb ischemia

• Published in: Biomaterials Vol. 34; no. 8; pp. 1995 - 2004
• Main Authors: Chen, Ding-Yuan, Wei, Hao-Ji, Lin, Kun-Ju, Huang, Chieh-Cheng, Wang, Chung-Chi, Wu, Cheng-Tse, Chao, Ko-Ting, Chen, Ko-Jie, Chang, Yen, Sung, Hsing-Wen
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Ltd 01.03.2013
• Subjects: Adhesion; Advanced Basic Science; Aggregates; Animals; Cell Aggregation - drug effects; Cell-based therapy; Collagen - pharmacology; Critical limb ischemia; Dentistry; Disease Models, Animal; Drug Combinations; Fetal Blood - cytology; Fluorescent Antibody Technique; Gene Expression Regulation - drug effects; Hindlimb - blood supply; Hindlimb - drug effects; Hindlimb - pathology; Human Umbilical Vein Endothelial Cells - cytology; Human Umbilical Vein Endothelial Cells - drug effects; Human Umbilical Vein Endothelial Cells - transplantation; Humans; Ischemia; Ischemia - pathology; Ischemia - therapy; Laminin - pharmacology; Limb Salvage; Limbs; Mesenchymal Stem Cell Transplantation; Mesenchymal Stromal Cells - cytology; Methylcellulose - chemistry; Mice; Mice, Inbred BALB C; Neovascularization; Neovascularization, Physiologic - drug effects; Networks; Perfusion; Proteoglycans - pharmacology; Regression; Regression analysis; Thermo-responsive hydrogel; Three dimensional; Tissue engineering; Veins; Cell-based therapy; Thermo-responsive hydrogel; Critical limb ischemia; Neovascularization; Tissue engineering
• ISSN: 0142-9612; 1878-5905
• DOI: 10.1016/j.biomaterials.2012.11.045

Abstract The proximity of cells in three-dimensional (3D) organization maximizes the cell–cell communication and signaling that are critical for cell function. In this study, 3D cell aggregates composed of human umbilical vein endothelial cells (HUVECs) and cord-blood mesenchymal stem cells (cbMSCs) were used for therapeutic neovascularization to rescue tissues from critical limb ischemia. Within the cell aggregates, homogeneously mixed HUVECs and cbMSCs had direct cell–cell contact with expressions of endogenous extracellular matrices and adhesion molecules. Although dissociated HUVECs/cbMSCs initially formed tubular structures on Matrigel, the grown tubular network substantially regressed over time. Conversely, 3D HUVEC/cbMSC aggregates seeded on Matrigel exhibited an extensive tubular network that continued to expand without regression. Immunostaining experiments show that, by differentiating into smooth muscle cell (SMC) lineages, the cbMSCs stabilize the HUVEC-derived tubular network. The real-time PCR analysis results suggest that, through myocardin, TGF-β signaling regulates the differentiation of cbMSCs into SMCs. Transplantation of 3D HUVEC/cbMSC aggregates recovered blood perfusion in a mouse model of hindlimb ischemia more effectively compared to their dissociated counterparts. The experimental results confirm that the transplanted 3D HUVEC/cbMSC aggregates enhanced functional vessel formation within the ischemic limb and protected it from degeneration. The 3D HUVEC/cbMSC aggregates can therefore facilitate the cell-based therapeutic strategies for modulating postnatal neovascularization.