title: A tag-less method for direct isolation of human umbilical vein endothelial cells by gravitational field-flow fractionation.
abstract: The analysis of cellular and molecular profiles represents a powerful tool in many biomedical applications to identify the mechanisms underlying the pathological changes. The improvement of cellular starting material and the maintenance of the physiological status in the sample preparation are very useful. Human umbilical vein endothelial cells (HUVEC) are a model for prediction of endothelial dysfunction. HUVEC are enzymatically removed from the umbilical vein by collagenase. This method provides obtaining a good sample yield. However, the obtained cells are often contaminated with blood cells and fibroblasts. Methods based on negative selection by in vitro passages or on the use of defined marker are currently employed to isolate target cells. However, these approaches cannot reproduce physiological status and they require expensive instrumentation. Here we proposed a new method for an easy, tag-less and direct isolation of HUVEC from raw umbilical cord sample based on the gravitational field-flow fractionation (GrFFF). This is a low-cost, fully biocompatible method with low instrumental and training investments for flow-assisted cell fractionation. The method allows obtaining pure cells without cell culture procedures as starting material for further analysis; for example, a proper amount of RNA can be extracted. The approach can be easily integrated into clinical and biomedical procedures.
authors.affiliation: Department of Obstetrics and Gynaecology, Fondazione IRCCS Cà Granda, Ospedale Maggiore Policlinico, Milan, Italy.
authors.forename: Debora
authors.initials: D
authors.lastname: Lattuada
authors.forename: Barbara
authors.initials: B
authors.lastname: Roda
authors.forename: Chiara
authors.initials: C
authors.lastname: Pignatari
authors.forename: Ruben
authors.initials: R
authors.lastname: Magni
authors.forename: Federico
authors.initials: F
authors.lastname: Colombo
authors.forename: Alessandra
authors.initials: A
authors.lastname: Cattaneo
authors.forename: Andrea
authors.initials: A
authors.lastname: Zattoni
authors.forename: Irene
authors.initials: I
authors.lastname: Cetin
authors.forename: Pierluigi
authors.initials: P
authors.lastname: Reschiglian
authors.forename: Giorgio
authors.initials: G
authors.lastname: Bolis
mesh_terms: D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D005260:Female; D031521:Fractionation, Field Flow / Q000295:instrumentation / Q000379:methods*; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology*; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D014470:Umbilical Cord / Q000166:cytology