Engineered Living Blood Vessels: Functional Endothelia Generated From Human Umbilical Cord-Derived Progenitors Dörthe Schmidt, MD, Lars M. Asmis, MD,

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Engineered Living Blood Vessels: Functional Endothelia Generated From Human Umbilical Cord-Derived Progenitors Dörthe Schmidt, MD, Lars M. Asmis, MD, Bernhard Odermatt, MD, Jens Kelm, PhD, Christian Breymann, MD, Matthias Gössi, PhD, Michele Genoni, MD, Gregor Zund, MD, Simon P. Hoerstrup, MD, PhD The Annals of Thoracic Surgery Volume 82, Issue 4, Pages 1465-1471 (October 2006) DOI: 10.1016/j.athoracsur.2006.05.066 Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 Macroscopic appearance of tissue-engineered blood vessels. Vascular scaffolds of 0.5 cm diameter and 3 cm length fabricated from nonwoven polyglycolic acid mesh and poly-4-hydroxybutyric acid before seeding (A) and after in vitro culturing (B). Tissue-engineered blood vessels were intact, pliable, and densely covered with cells (B). The lumen was open, and the wall thickness was homogeneous at about 0.3 cm. The Annals of Thoracic Surgery 2006 82, 1465-1471DOI: (10.1016/j.athoracsur.2006.05.066) Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 Histology and immunohistochemistry of tissue-engineered blood vessels. (A) Hematoxylin and eosin staining revealed a cellular tissue with an outer layer consisting of loosely arranged cells and a middle layer organized more compactly. The innermost layer consisted of endothelial cells. (B) Cells in the middle layer expressed α-smooth muscle actin. (C) A CD31-positive cell layer lined the lumen of the tissue-engineered blood vessels. Additionally, some cells expressing CD31 could be found in the middle layer. (D) Higher magnification of CD31-positive cells distant from the luminal cell lining. Arrows point at remnants of scaffold. The Annals of Thoracic Surgery 2006 82, 1465-1471DOI: (10.1016/j.athoracsur.2006.05.066) Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Morphology of the inner surfaces. Scanning electron micrographs show smooth inner surfaces of the tissue-engineered blood vessels densely covered with endothelial cells derived from endothelial progenitor cells (A) and human umbilical vein endothelial cells (B). The Annals of Thoracic Surgery 2006 82, 1465-1471DOI: (10.1016/j.athoracsur.2006.05.066) Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 Quantification of extracellular matrix elements and cell number. Total cell number (represented as DNA) in tissue-engineered blood vessels was 90% of native tissue values; glycosaminoglycans (GAG) and collagen (hydroxyproline, HYP) were 62% and 1%, respectively. The Annals of Thoracic Surgery 2006 82, 1465-1471DOI: (10.1016/j.athoracsur.2006.05.066) Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions

Fig 5 Functional plasticity and bioactivity of endothelial cells. Both endothelial progenitor cell–derived endothelial cells (EPC) and human umbilical vein endothelial cells (HUVEC) expressed CD31 and thrombomodulin (TM) but lacked the expression of tissue factor (TF) (grey). After stimulation with tumor necrosis factor-α (black) the expression of thrombomodulin was decreased whereas the expression of tissue factor was upregulated. CD31 expression was not affected. The Annals of Thoracic Surgery 2006 82, 1465-1471DOI: (10.1016/j.athoracsur.2006.05.066) Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions