Origin of endothelial progenitors in human postnatal bone marrow

Abstract

MethodsMAPC cultures.BM was obtained from 55 healthy volunteer donors (2-45 years of age) after obtaining informed consent per the guidelines of the University of Minnesota Committee on the Use of Human Subjects in Research.MAPCs were generated as previously described (3).Briefly, BM mononuclear cells were depleted of CD45 + and glycophorin A + cells using micromagnetic beads (Miltenyi Biotec, Sunnyvale, California, USA).Cells (5 10 3 ) that were negative for CD45 and glycophorin A were diluted in 200 l expansion medium consisting of 58% low-glucose DMEM (Invitrogen Corp., Grand Island, New York, USA) and 40% MCDB-201 (Sigma Chemical Co., St. Louis, Missouri, USA), supplemented with 1 insulin-transferrinselenium, 1 linoleic acid-BSA, 10 -8 M dexamethasone, 10 -4 M ascorbic acid 2-phosphate (all from Sigma Chemical Co.), 100 U penicillin, and 1,000 U streptomycin (Invitrogen Corp.); along with 0-10% FCS (HyClone Laboratories, Logan, Utah, USA), 10 ng/ml EGF (Sigma Chemical Co.), and 10 ng/ml PDGF-BB