title: Neovascularization Potential of Blood Outgrowth Endothelial Cells From Patients With Stable Ischemic Heart Failure Is Preserved.
abstract: BACKGROUND
  Blood outgrowth endothelial cells (BOECs) mediate therapeutic neovascularization in experimental models, but outgrowth characteristics and functionality of BOECs from patients with ischemic cardiomyopathy (ICMP) are unknown. We compared outgrowth efficiency and in vitro and in vivo functionality of BOECs derived from ICMP with BOECs from age-matched (ACON) and healthy young (CON) controls.
  
  
  RESULTS
  We isolated 3.6±0.6 BOEC colonies/100×10(6) mononuclear cells (MNCs) from 60-mL blood samples of ICMP patients (n=45; age: 66±1 years; LVEF: 31±2%) versus 3.5±0.9 colonies/100×10(6) MNCs in ACON (n=32; age: 60±1 years) and 2.6±0.4 colonies/100×10(6) MNCs in CON (n=55; age: 34±1 years), P=0.29. Endothelial lineage (VEGFR2(+)/CD31(+)/CD146(+)) and progenitor (CD34(+)/CD133(-)) marker expression was comparable in ICMP and CON. Growth kinetics were similar between groups (P=0.38) and not affected by left ventricular systolic dysfunction, maladaptive remodeling, or presence of cardiovascular risk factors in ICMP patients. In vitro neovascularization potential, assessed by network remodeling on Matrigel and three-dimensional spheroid sprouting, did not differ in ICMP from (A)CON. Secretome analysis showed a marked proangiogenic profile, with highest release of angiopoietin-2 (1.4±0.3×10(5) pg/10(6) ICMP-BOECs) and placental growth factor (5.8±1.5×10(3) pg/10(6) ICMP BOECs), independent of age or ischemic disease. Senescence-associated β-galactosidase staining showed comparable senescence in BOECs from ICMP (5.8±2.1%; n=17), ACON (3.9±1.1%; n=7), and CON (9.0±2.8%; n=13), P=0.19. High-resolution microcomputed tomography analysis in the ischemic hindlimb of nude mice confirmed increased arteriogenesis in the thigh region after intramuscular injections of BOECs from ICMP (P=0.025; n=8) and CON (P=0.048; n=5) over vehicle control (n=8), both to a similar extent (P=0.831).
  
  
  CONCLUSIONS
  BOECs can be successfully culture-expanded from patients with ICMP. In contrast to impaired functionality of ICMP-derived bone marrow MNCs, BOECs retain a robust proangiogenic profile, both in vitro and in vivo, with therapeutic potential for targeting ischemic disease.
authors.affiliation: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium dieter.dauwe@med.kuleuven.be stefan.janssens@med.kuleuven.be.
authors.forename: Dieter
authors.initials: D
authors.lastname: Dauwe
authors.affiliation: Cell Therapy Department, Center for Applied Medicine Research, Clinica Universidad de Navarra, University of Navarra, Pamplona, Spain.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Pelacho
authors.affiliation: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
authors.forename: Arief
authors.initials: A
authors.lastname: Wibowo
authors.affiliation: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
authors.forename: Ann-Sophie
authors.initials: AS
authors.lastname: Walravens
authors.affiliation: Center for Molecular and Vascular Biology, KU Leuven, Leuven, Belgium.
authors.forename: Kristoff
authors.initials: K
authors.lastname: Verdonck
authors.affiliation: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
authors.forename: Hilde
authors.initials: H
authors.lastname: Gillijns
authors.affiliation: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
authors.forename: Ellen
authors.initials: E
authors.lastname: Caluwe
authors.affiliation: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
authors.forename: Peter
authors.initials: P
authors.lastname: Pokreisz
authors.affiliation: Department of Clinical and Experimental Endocrinology, KU Leuven, Leuven, Belgium.
authors.forename: Nick
authors.initials: N
authors.lastname: van Gastel
authors.affiliation: Department of Clinical and Experimental Endocrinology, KU Leuven, Leuven, Belgium.
authors.forename: Geert
authors.initials: G
authors.lastname: Carmeliet
authors.affiliation: Department of Electrical Engineering, Center for the Processing of Speech and Images, KU Leuven, Leuven, Belgium.
authors.forename: Maarten
authors.initials: M
authors.lastname: Depypere
authors.affiliation: Department of Electrical Engineering, Center for the Processing of Speech and Images, KU Leuven, Leuven, Belgium.
authors.forename: Frederik
authors.initials: F
authors.lastname: Maes
authors.affiliation: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
authors.forename: Nina
authors.initials: N
authors.lastname: Vanden Driessche
authors.affiliation: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
authors.forename: Walter
authors.initials: W
authors.lastname: Droogne
authors.affiliation: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
authors.forename: Johan
authors.initials: J
authors.lastname: Van Cleemput
authors.affiliation: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
authors.forename: Johan
authors.initials: J
authors.lastname: Vanhaecke
authors.affiliation: Cell Therapy Department, Center for Applied Medicine Research, Clinica Universidad de Navarra, University of Navarra, Pamplona, Spain Hematology Department, Clinica Universidad de Navarra, University of Navarra, Pamplona, Spain.
authors.forename: Felipe
authors.initials: F
authors.lastname: Prosper
authors.affiliation: Department of Development and Regeneration, Stem Cell Biology and Embryology, KU Leuven, Leuven, Belgium.
authors.forename: Catherine
authors.initials: C
authors.lastname: Verfaillie
authors.affiliation: Center for Molecular and Vascular Biology, KU Leuven, Leuven, Belgium.
authors.forename: Aernout
authors.initials: A
authors.lastname: Luttun
authors.affiliation: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium dieter.dauwe@med.kuleuven.be stefan.janssens@med.kuleuven.be.
authors.forename: Stefan
authors.initials: S
authors.lastname: Janssens
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D016022:Case-Control Studies; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000503:physiopathology* / Q000637:transplantation; D005260:Female; D006801:Humans; D008297:Male; D008819:Mice, Nude; D008875:Middle Aged; D017202:Myocardial Ischemia / Q000503:physiopathology*; D018919:Neovascularization, Physiologic / Q000502:physiology*; D018384:Oxidative Stress / Q000502:physiology; D055815:Young Adult
keywords: arteriogenesis; blood outgrowth endothelial cells; cell transplantation; ischemic heart disease; therapeutic neovascularization