A Xeno-Free Strategy for Derivation of Human Umbilical Vein Endothelial Cells and Wharton’s Jelly Derived Mesenchymal Stromal Cells: A Feasibility Study toward Personal Cell and Vascular Based Therapy

Abstract

Coimplantation of endothelial cells (ECs) and mesenchymal stromal cells (MSCs) into the transplantation site could be a feasible option to achieve a sufficient level of graft-host vascularization. To find a suitable source of tissue that provides a large number of high-quality ECs and MSCs suited for future clinical application, we developed a simplified xeno-free strategy for isolation of human umbilical vein endothelial cells (HUVECs) and Wharton’s jelly-derived mesenchymal stromal cells (WJ-MSCs) from the same umbilical cord. We also assessed whether the coculture of HUVECs and WJ-MSCs derived from the same umbilical cord (autogenic cell source) or from different umbilical cords (allogenic cell sources) had an impact on in vitro angiogenic capacity. We found that HUVECs grown in 5 ng/ml epidermal growth factor (EGF) supplemented xeno-free condition showed higher proliferation potential compared to other conditions. HUVECs and WJ-MSCs obtained from this technic show an endothelial lineage (CD31 and von Willebrand factor) and MSC (CD73, CD90, and CD105) immunophenotype characteristic with high purity, respectively. It was also found that only the coculture of HUVEC/WJ-MSC, but not HUVEC or WJ-MSC mono-culture, provides a positive effect on vessel-like structure (VLS) formation, in vitro. Further investigations are needed to clarify the pros and cons of using autogenic or allogenic source of EC/MSC in tissue engineering applications. To the best of our knowledge, this study offers a simple, but reliable, xeno-free strategy to establish ECs and MSCs from the same umbilical cord, a new opportunity to facilitate the development of personal cell-based therapy.

Co-culture of liver parenchymal cells with non-parenchymal cells under 2D and 3D culture systems; a review paper

“…HUVEC has easy isolation methods without contamination with other cells [77]. Expression of the vWF marker distinct these cells from fibroblast or smooth muscle cells [78].…”

Applications of extraembryonic tissue-derived cells in vascular tissue regeneration

“…These effects involve stabilizing the EC network and secreting vasculogenic growth factors, including hepatocyte growth factor. In both autogenic and allogenic cell sources, WJ-MSCs have been shown to promote angiogenesis and enhance vascular tube formation when cocultured with ECs for three days [ 81 ].…”

“… [ 92 ] 10 HUVECs WJ-MSCs The coculture of HUVECs and WJMSC can develop VLS after 3 days in an endothelial culture medium. [ 81 ] 11 WJ-MSCs chitosan/hyaluronic acid WJ-MSCs differentiated into endothelial-like cells on chitosan/hyaluronic acid (HA/CHI) multilayers. [ 76 ] 12 WJ-MSCs decellularized hUAs The cord blood platelet lysate improved the repopulation efficacy.…”

Implications of Endothelial Cell-Mediated Dysfunctions in Vasomotor Tone Regulation

“…Over time, these studies have undergone some adaptations, such as the constitution of the culture medium for endothelial cells. In this sense, the constitution of the culture medium used may contain endothelial cell growth factor (ECGF), which allows long-term growth of the ECs, or an anti-PPLO agent that prevents contamination by gram-positive bacteria or mycoplasmas [41,[44][45][46][47]. Culture media containing 5 ng/mL epidermal growth factor (EGF) was shown to have a higher proliferative potential compared to the anti-pleuropneumonia-like organisms' agent (anti-PPLO agent).…”