Isolation and longterm culture of human intestinal microvascular endothelial cels

G. Haraldsen*, J. Rugtveit, D. Kvale, T. Scholz, W. A. Muller, T. Hovig, P. Brandtzaeg

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

Abstract

Microvascular endothelial cells play an important part in inflammation as well as in organ specific leucocyte traffic, and may be functionally different from large vessel endothelium in this respect. This study therefore established a method for isolation and longterm culture of human intestinal microvascular endothelial cells (HIMEC). After dissociation by collagenase/dispase/DNase of mucosal and submucosal tissue obtained from normal adult jejunum, cells were plated and cultured to subconfluence in endothelial serum free medium containing 2.5% fetal calf serum, hydrocortisone, and N6, O2-dibutyryladenosine cyclic monophosphate. Primary cultures were trypsinised and endothelial cells were isolated by paramagnetic beads armed with monoclonal antibody to CD31. Optimal growth conditions for HIMEC cultures were established, allowing up to nine passages (three months in vitro). The cells contained Weibel-Palade bodies, expressed von Willebrand factor, CD31, and VE-cadherin; and bound Ulex Europaeus lectin I. A method to establish longterm cell cultures of HIMEC will facilitate further investigation of the function of intestinal endothelial cells and their participation in physiological and pathological events in the gut.

Original languageEnglish (US)
Pages (from-to)225-234
Number of pages10
JournalGut
Volume37
Issue number2
DOIs
StatePublished - 1995

Keywords

  • cell separation
  • cyclic AMP
  • intestinal mucosa
  • microcirculation
  • vascular endothelium

ASJC Scopus subject areas

  • Gastroenterology

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