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Analytical and Bioanalytical Chemistry

, Volume 405, Issue 2–3, pp 977–984 | Cite as

A tag-less method for direct isolation of human umbilical vein endothelial cells by gravitational field-flow fractionation

  • Debora Lattuada
  • Barbara RodaEmail author
  • Chiara Pignatari
  • Ruben Magni
  • Federico Colombo
  • Alessandra Cattaneo
  • Andrea Zattoni
  • Irene Cetin
  • Pierluigi Reschiglian
  • Giorgio Bolis
Original Paper

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.

Keywords

Tag-less cell sorting Gravitational field-flow fractionation (GrFFF) HUVEC (Human umbilical vein endothelial cells) 

Notes

Acknowledgements

We are grateful to Chiara Bolego for her precious help and suggestions in HUVEC culture and their isolation from umbilical cord sample.

Competing interest

Barbara Roda, Andrea Zattoni, and Pierluigi Reschiglian are associates of the academic spinoff company byFlow Srl (Bologna, Italy). The company mission includes know-how transfer, development, and application of novel technologies and methodologies for the analysis and characterization of samples of nano-biotechnological interest.

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Debora Lattuada
    • 1
  • Barbara Roda
    • 2
    • 5
    Email author
  • Chiara Pignatari
    • 1
  • Ruben Magni
    • 1
  • Federico Colombo
    • 3
  • Alessandra Cattaneo
    • 3
  • Andrea Zattoni
    • 2
    • 5
  • Irene Cetin
    • 4
  • Pierluigi Reschiglian
    • 2
    • 5
  • Giorgio Bolis
    • 6
  1. 1.Department of Obstetrics and GynaecologyFondazione IRCCS Cà Granda, Ospedale Maggiore PoliclinicoMilanItaly
  2. 2.Department of Chemistry “G. Ciamician”BolognaItaly
  3. 3.Centre of Transfusion Medicine, Cellular Therapy and CryobiologyFoundation IRCCS Ca’ Granda, Ospedale Maggiore PoliclinicoMilanItaly
  4. 4.Unit of Obstetrics and Gynecology, Department of Clinical Sciences L. SaccoUniversity of MilanMilanItaly
  5. 5.byFlow SrlBolognaItaly
  6. 6.Department of Mother and Infant Sciences, Foundation IRCCS Ca’ Granda, Ospedale Maggiore PoliclinicoUniversity of MilanMilanItaly

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