A tag-less method for the direct isolation of HUVEC (human umbilical vein endothelial cells) by gravitational field-flow fractionation as a starting point for gene expression analysis

HUVECs (human umbilical vein endothelial cells) are a model for prediction of endothelial dysfunction in various diseases. Cultured cells seem to be a good model for studying molecular processes in endothelial tissue. Nevertheless, they cannot reproduce real physiological status because some biological factors necessarily change in culture conditions. As a consequence, it would be useful to improve preparation of the cellular starting materials from real samples, especially when gene expression studies are further required on the cells. In this work we present a new method for the easy isolation of HUVECs from raw cord blood based on the use of the flow-assisted separative technique, gravitational field-flow fractionation (GrFFF) able to give a relatively pure sample for gene analysis without cell culture steps. The method can be applied to cell populations enzymatically removed from the umbilical vein which can be purified from contaminants cells and used directly for gene expression analysis.