OC48 MYOCARDIAL TISSUE ENGINEERING AND REGENERATION– IN VITRO EXPANSION OF ENDOTHELIAL CELL BY DIFFERENT VESSELS REVEALS THE BETTER AUTOLOGOUS CELLULAR SOURCE FOR TISSUE REPLACEMENT

Abstract

BACKGROUND AND AIM: The development of cell-based therapies to assist/replace diseased tissue is one of the most promising initiatives in regenerative medicine. Human endothelial cells (HEC) can be used for various applications including molecular profiling as well as cell culture. Several complex techniques and culture media have been reported although with confounding results belonging todonor-to-donor variability and cellular source of derivation. Our results demonstrate the overcome of these limitations using soluble CD54 (sCD54) as additive to conventional culture medium. METHODS: Isolated primary fragment of different vessel types (left internal thoracic artery, radial artery and saphenous vein) were expanded in Ham's F12 DMEM, enriched with growth factors, Foetal Calf Serum and conditioned medium of Human Umbilical Vein Endothelial Cells (HUVEC) collected at different passages. Cytokines were analysed in order to identify the soluble factors correlating with better proliferation profile. RESULTS: sCD54 was found to induce the in vitro expansion of HEC independently from the vessels source and even in the absence of HUVEC-conditioned medium. The HECs cultivated in presence of sCD54 resulted positive for the expression of CD146 and negative for CD45, and with lower fibroblast contamination. Cells were capable to proliferate with an S phase of 25%, to produce VEGF and to give origin to vessel-like tubule in vitro. (Figure 1) CONCLUSIONS: sCD54 is an essential factor for the in-vitro expansion and differentiation of HEC, minimizing potential donor sample variability. Resulting primary cultures can be advantageous for tissue engineering in regenerative medicine and bio-nanotechnology applications.