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.

Rangsun Parnpai,Tanut Kunkanjanawan,Rungrueang Yodsheewan,Veerapol Khemarangsan,Kasem Theerakittayakorn,Hataiwan Kunkanjanawan

doi:10.1155/2020/8832052

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.

Highlights

Since blood supply is an essential factor that holds the great effect on graft survival and host tissue integration, various approaches promoting vascularization have been developed in the field of tissue engineering research
To creating a new opportunity to facilitate the development of personal cell and vascular-based therapy, the objectives of this study are to isolate and expand human umbilical vein endothelial cells (HUVECs) and WJ-mesenchymal stromal cells (MSCs) from the same umbilical cord using the defined xeno-free strategies and to determine how the coculture of autogenic and allogenic HUVEC/Wharton’s jelly-derived mesenchymal stromal cells (WJ-MSCs) contribute to the angiogenic capacity, in vitro
To obtain multiple cell types potentially useful for cell and vascular based therapy, we first examined the possibility of isolation of HUVECs and WJMSCs from single umbilical cord using a well-defined xenofree culture system compared with commercially available xeno-containing culture medium

Summary

Introduction

Since blood supply is an essential factor that holds the great effect on graft survival and host tissue integration, various approaches promoting vascularization have been developed in the field of tissue engineering research. Cotransplantation of multiple cell types (i.e., adiposederived stem cells and human umbilical vein endothelial cells; HUVECs) has proven to yield a superiority effect on neovascularization in an adipogenesis mouse models [1]. This finding is in accordance with Ma et al (2014), who showed that coculture of human adipose tissue-derived or. Several groups have reported various protocols for the isolation of Wharton’s jelly-derived mesenchymal stromal cells (WJ-MSCs) from hUC using animal-free or socalled xeno-free culture system [7,8,9,10]. To comply with the long-term safety requirements for cell-based therapy, xenofree established cells have become a preferred source of cellbased product suited for future clinical application [15]

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