Authors: D. Lattuada, B. Roda, C. Pignatari, R. Magni, A. Cattaneo, G. Bolis, P. Reschiglian
Affilation: University of Bologna, Italy
Pages: 191 - 194
Keywords: cell separation, gravitational field-flow fractionation, gene expression analysis
HUVECs (human umbilical vein endothelial cells) are a model for prediction of endothelial dysfunction in various diseases. HUVECs are enzymatically removed from the umbilical vein by collagenase. This method allows for a good sample yield, but so-obtained HUVECs are contaminated with red blood cells (RBCs) and fibroblasts, which need to be negatively selected by in vitro passages. 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. GrFFF is a low-cost, fully biocompatible, tag-less method for flow-assisted cell fractionation. The channel is empty, the fractionation mechanism is soft and it doesn’t modify the native properties of cells. Cell suspension are injected into the GrFFF device and in relatively short time (10 minutes) separation is archived. Due to the high reproducibility of fractionation method, cells from different injection can be cumulated in order to obtain a cell preparation suitable for gene expression analysis. Post separation samples had purity higher than 90%. Obtaining a good amount of purified cells could avoid cell cultures and be useful for further studies on cell.