ISOLATION AND LONG-TERM CULTURE OF HUMAN INTESTINAL MICROVASCULAR ENDOTHELIAL-CELLS

Microvascular endothelial cells play an important part in inflammation as well as in organ specific leucocyte traffic, and may be functional ly 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 dissoc iation by collagenase/dispase/DNase of mucosal and submucosal tissue o btained from normal adult jejunum, cells were plated and cultured to s ubconfluence in endothelial serum free medium containing 2.5% fetal ca lf serum, hydrocortisone, and N-6, O-2-dibutyryladenosine cyclic monop hosphate. 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, allow ing up to nine passages (three months in vitro). The cells contained W eibel-Palade bodies, expressed von Willebrand factor, CD31, and VE-cad herin; and bound Ulex Europaeus lectin I. A method to establish longte rm cell cultures of HIMEC will facilitate further investigation of the function of intestinal endothelial cells and their participation in p hysiological and pathological events in the gut.