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Phenotypic differences in putative EPCs based on in vitro cultivation (855.8)
Author(s) -
Rocha Natalia,
Bammert Tyler,
Beckstrom Collin,
Kavlich Philip,
Lincenberg Grace,
Greiner Jared,
DeSouza Christopher
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.855.8
Subject(s) - peripheral blood mononuclear cell , progenitor cell , ficoll , population , fibronectin , phenotype , hepatocyte growth factor , differential centrifugation , biology , andrology , apoptosis , microbiology and biotechnology , in vitro , immunology , chemistry , stem cell , medicine , extracellular matrix , genetics , gene , receptor , environmental health
Circulating endothelial progenitor cells (EPCs) have been extensively studied over the past decade. Isolation techniques have differed in the literature based on whether the non‐adherent or adherent cells from fibronectin‐coated plates are studied. We determined whether there are phenotypic differences in cellular function between putative EPCs derived from the adherent or non‐adherent mononuclear cell population. Peripheral blood samples were collected from 15 healthy adults. Mononuclear cells were isolated by Ficoll density‐gradient centrifugation and plated on 6‐well plates coated with human fibronectin. After 2 days, non‐adherent cells were collected; in a separate culture, adherent cells were removed and collected after 7 days. Migratory capacity (Boyden chamber), angiogenic growth factor release (ELISA) and apoptosis (TUNEL) were assessed in both cell types. Migration to both VEGF (505+86 vs 254+ 55 AU) and SDF‐1 (498+83 vs 231+46 AU) were ~2‐fold higher (P<0.05) in the non‐adherent vs adherent cells. Release of angiogenic factors: granulocyte colony‐stimulating factor and hepatocyte growth factor were not different between cell types. Apoptotic response to staurosporine was significantly lower (~60%) in non‐adherent cells. In summary, non‐adherent and adherent cultured putative EPCs demonstrate functional phenotypic differences in migratory capacity and apoptotic susceptibility.