Production of Human Endothelial Cells Free from Soluble Xenogeneic Antigens for Bioartificial Small Diameter Vascular Graft Endothelization

Abstract

Arterial bypass graft implantation remains the primary therapy for patients with advanced cardiovascular disease, but most lack adequate saphenous vein or other conduits for bypass procedures and would benefit from a bioartificial conduit. This study aimed to produce human endothelial cells (hECs) in large scale, free from xenogeneic antigens, to develop a small diameter, compatible vessel for potential use as a vascular graft. Human adipose-derived stromal cells (hASCs) were isolated, cultured, and differentiated in the presence of human serum and used for the reendothelization of a decellularized rat aorta. hASC derived ECs (hASC-ECs) expressed VEGFR2, vWf and CD31 endothelial cell markers, the latter in higher levels than hASCs and HUVECs, and were shown to be functional. Decellularization protocol yielded aortas devoid of cell nuclei, with preserved structure, including a preserved basement membrane. When seeded with hASC-ECs, the decellularized aorta was completely reendothelized, and the hASC-ECs maintained their phenotype in this new condition. hASCs can be differentiated into functional hECs without the use of animal supplements and are capable of reendothelizing a decellularized rat aorta while maintaining their phenotype. The preservation of the basement membrane following decellularization supported the complete reendothelization of the scaffold with no cell migration towards other layers. This approach is potentially useful for rapid obtention of compatible, xenogeneic-free conduit.

A Scalable and Efficient Bioprocess for Manufacturing Human Pluripotent Stem Cell-Derived Endothelial Cells

“…They are of great value for disease modeling, drug screening, cell therapy, and tissue engineering ( Heo et al., 2014 , Huang et al., 2013 , Kang et al., 2013 , Leung et al., 2016 , Medina et al., 2010 , Moubarik et al., 2011 , Patsch et al., 2015 , Schwarz et al., 2012 , Stroncek et al., 2012 ). However, obtaining large numbers of primary ECs for those applications, in particular for clinical applications ( Arici et al., 2015 , Chao et al., 2014 , den Dekker et al., 2011 , Granton et al., 2015 , Matoba et al., 2008 ), is still challenging due to their limited proliferation capacity and phenotype changes during the in vitro culture ( van Beijnum et al., 2008 , de Carvalho et al., 2015 , Gui et al., 2009 , Gumbleton and Audus, 2001 , Hayflick, 1965 , Augustin-Voss et al., 1993 ). Human pluripotent stem cells (hPSCs) provide a potential solution to this challenge ( Levenberg et al., 2007 ).…”

Applied Bioengineering in Tissue Reconstruction, Replacement, and Regeneration

“…In recent years, the focus of this research has been finding more biocompatible and biodegradable matrices to seed with ECs. Endothelial and SMCs utilized in seeding cellularized grafts have several potential sources, including harvesting of endothelial progenitor cells isolated from peripheral blood, bone marrow, adipose tissue, umbilical cord blood, or blood vessel walls for ex vivo expansion, 258,259 endothelial and SMCs derived from human iPSCs, 260 ECs derived from hASCs, 261 or autologous cells obtained through biopsy. These cells are cultivated in vitro under specific conditions and characterized by endothelialspecific morphology and biomarkers.…”

Comparative Study of Human Pluripotent Stem Cell-Derived Endothelial Cells in Hydrogel-Based Culture Systems

“… 3 − 11 However, because of the limited proliferation capability and quick phenotype change during in vitro culturing, obtaining enough primary ECs for basic applications remains very challenging. 12 − 17 Human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) 18 and induced pluripotent stem cells (iPSCs), 19 , 20 provide a potential solution to this challenge 21 due to their unlimited proliferation capability and the ability to differentiate into all somatic cell types of the human body. 22 , 23 In particular, patient-derived iPSCs contain the patient’s genetic information and could model many human diseases.…”

“…Endothelial cells (ECs), which play an important role in normal vascular functions and a variety of vascular diseases, , are promising cell sources for drug screening, cell therapy, and tissue engineering. − However, because of the limited proliferation capability and quick phenotype change during in vitro culturing, obtaining enough primary ECs for basic applications remains very challenging. − Human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs), , provide a potential solution to this challenge due to their unlimited proliferation capability and the ability to differentiate into all somatic cell types of the human body. , In particular, patient-derived iPSCs contain the patient’s genetic information and could model many human diseases. Currently, the methods of hPSC differentiation into ECs in a 3D suspension ,− or 2D monolayer ,− have been established.…”