The isolation and culture of endothelial colony-forming cells from human and rat lungs

Abstract

Blood vessels are crucial for the normal development, lifelong repair and homeostasis of tissues.Recently, vascular progenitor cell-driven "postnatal vasculogenesis" has been suggested as an important mechanism contributing to new blood vessel formation and organ repair. Among several described progenitor cell types contributing to blood vessel formation, endothelial colony forming cells (ECFCs) have received widespread attention as lineage-specific 'true' vascular progenitors. Herein, we describe a protocol for the isolation of pulmonary microvascular ECFCs from human and rat lung tissue. Our technique takes advantage of an earlier protocol for isolating circulating ECFCs from the mononuclear cellular fraction of peripheral blood. We

Concise Review: Functional Definition of Endothelial Progenitor Cells: A Molecular Perspective

“…Although this is a widely used functional assay, many cell types independent of endothelial origin can also spontaneously form tubes, and therefore this assay lacks specificity and must be combined with other tangible markers of differentiation [38]. Surface expression of lectins, endocytosis of acetylated low-density lipoprotein, expression of endothelial specific adhesion, and function genes are more or less systematically proposed to fill this shortcoming [39].…”

Homing of Cultured Endothelial Progenitor Cells and Their Effect on Traumatic Brain Injury in Rat Model

“…After culturing for three days, floating cells were removed by washing with potassium-buffered saline (PBS), and new media was added to the adherent cells. The cells were maintained in culture for seven days 2, 3 .…”

“…After washing with PBS, FITC-labeled lectin from ulexeuropaeus agglutinin (UEA) (Biomeda) was added to the cells for 1 h. Samples were washed with PBS, and viewed under an inverted fluorescent microscope and a laser scanning confocal microscope (Leica, Germany). Cells exhibiting double-positive fluorescence were identified as differentiating EPCs 2, 3 .…”

Vasculogenesis and Its Cellular Therapeutic Applications

“…Another source of tissue EPCs is the microvessels of the lung, placenta and white adipose tissue [Lin et al, 2013;Patel et al, 2013;Alphonse et al, 2014Alphonse et al, , 2015. Rat Ratajska et al Cells Tissues Organs 2017;203:141-152 DOI: 10.1159/000448551 146 (and human) pulmonary microvascular ECs contain a subset of CD31 + cells of high proliferative and vasculogenic activity which form perfused vessels when implanted on Matrigel plugs in vivo [Schniedermann et al, 2010;Basile and Yoder, 2014;Alphonse et al, 2015].…”

“…Rat Ratajska et al Cells Tissues Organs 2017;203:141-152 DOI: 10.1159/000448551 146 (and human) pulmonary microvascular ECs contain a subset of CD31 + cells of high proliferative and vasculogenic activity which form perfused vessels when implanted on Matrigel plugs in vivo [Schniedermann et al, 2010;Basile and Yoder, 2014;Alphonse et al, 2015]. These cells are more numerous than the EPCs found in the rat pulmonary artery [Lin et al, 2013;Patel et al, 2013;Alphonse et al, 2014Alphonse et al, , 2015Basile and Yoder, 2014]. It is assumed that EPCs are easily mobilized from the BM or the CB to these niche locations in solid tissues [Hirschi et al, 2008].…”