title: Increased yield of endothelial cells from peripheral blood for cell therapies and tissue engineering.
abstract: OBJECTIVE
  Peripheral blood-derived endothelial cells (pBD-ECs) are an attractive tool for cell therapies and tissue engineering, but have been limited by their low isolation yield. We increase pBD-EC yield via administration of the chemokine receptor type 4 antagonist AMD3100, as well as via a diluted whole blood incubation (DWBI).
  
  
  METHODS
  Porcine pBD-ECs were isolated using AMD3100 and DWBI and tested for EC markers, acetylated LDL uptake, growth kinetics, metabolic activity, flow-mediated nitric oxide production and seeded onto titanium tubes implanted into vessels of pigs.
  
  
  RESULTS
  DWBI increased the yield of porcine pBD-ECs 6.6-fold, and AMD3100 increased the yield 4.5-fold. AMD3100-mobilized ECs were phenotypically indistinguishable from nonmobilized ECs. In porcine implants, the cells expressed endothelial nitric oxide synthase, reduced thrombin-antithrombin complex systemically and prevented thrombosis.
  
  
  CONCLUSIONS
  Administration of AMD3100 and the DWBI method both increase pBD-EC yield.
authors.affiliation: 1Department of Surgery, Duke University Medical Center, NC, USA.
authors.forename: Ryan M
authors.initials: RM
authors.lastname: Jamiolkowski
authors.affiliation: 1Department of Surgery, Duke University Medical Center, NC, USA.
authors.forename: Sa Do
authors.initials: SD
authors.lastname: Kang
authors.affiliation: 1Department of Surgery, Duke University Medical Center, NC, USA.
authors.forename: AnnMarie K
authors.initials: AK
authors.lastname: Rodriguez
authors.affiliation: 1Department of Surgery, Duke University Medical Center, NC, USA.|2Department of Biomedical Engineering, Duke University, NC, USA.
authors.forename: Justin M
authors.initials: JM
authors.lastname: Haseltine
authors.affiliation: 1Department of Surgery, Duke University Medical Center, NC, USA.|2Department of Biomedical Engineering, Duke University, NC, USA.
authors.forename: Lauren J
authors.initials: LJ
authors.lastname: Galinat
authors.affiliation: 2Department of Biomedical Engineering, Duke University, NC, USA.
authors.forename: Alexandra E
authors.initials: AE
authors.lastname: Jantzen
authors.affiliation: 1Department of Surgery, Duke University Medical Center, NC, USA.|2Department of Biomedical Engineering, Duke University, NC, USA.
authors.forename: Tim A
authors.initials: TA
authors.lastname: Carlon
authors.affiliation: 1Department of Surgery, Duke University Medical Center, NC, USA.
authors.forename: Marcus D
authors.initials: MD
authors.lastname: Darrabie
authors.affiliation: 1Department of Surgery, Duke University Medical Center, NC, USA.
authors.forename: Antonio J
authors.initials: AJ
authors.lastname: Arciniegas
authors.affiliation: 1Department of Surgery, Duke University Medical Center, NC, USA.
authors.forename: Jose G
authors.initials: JG
authors.lastname: Mantilla
authors.affiliation: 3Duke Translational Medicine Institute, NC, USA.
authors.forename: N Rebecca
authors.initials: NR
authors.lastname: Haley
authors.affiliation: 1Department of Surgery, Duke University Medical Center, NC, USA.|4Duke-National University of Singapore Graduate Medical School, Singapore.
authors.forename: Maria
authors.initials: M
authors.lastname: Noviani
authors.affiliation: 5Department of Community and Family Medicine, Duke University Medical Center, NC, USA.|6Duke Molecular Physiology Institute, Duke University Medical Center, NC, USA.
authors.forename: Jason D
authors.initials: JD
authors.lastname: Allen
authors.affiliation: 6Duke Molecular Physiology Institute, Duke University Medical Center, NC, USA.
authors.forename: Thomas V
authors.initials: TV
authors.lastname: Stabler
authors.affiliation: 1Department of Surgery, Duke University Medical Center, NC, USA.
authors.forename: James W
authors.initials: JW
authors.lastname: Frederiksen
authors.affiliation: 8University of North Carolina, School of Medicine, NC, USA.
authors.forename: Oscar
authors.initials: O
authors.lastname: Alzate
authors.affiliation: 8University of North Carolina, School of Medicine, NC, USA.
authors.forename: Lukas G
authors.initials: LG
authors.lastname: Keil
authors.affiliation: 8University of North Carolina, School of Medicine, NC, USA.
authors.forename: Siyao
authors.initials: S
authors.lastname: Liu
authors.affiliation: 1Department of Surgery, Duke University Medical Center, NC, USA.
authors.forename: Fu-Hsiung
authors.initials: FH
authors.lastname: Lin
authors.affiliation: 2Department of Biomedical Engineering, Duke University, NC, USA.
authors.forename: George A
authors.initials: GA
authors.lastname: Truskey
authors.affiliation: 1Department of Surgery, Duke University Medical Center, NC, USA.|9Department of Pathology, Duke University Medical Center, NC, USA.
authors.forename: Hardean E
authors.initials: HE
authors.lastname: Achneck
mesh_terms: D000818:Animals; D001596:Benzylamines; D002469:Cell Separation; D017690:Cell Transplantation / Q000379:methods*; D000080027:Cyclams; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects; D005434:Flow Cytometry; D006571:Heterocyclic Compounds / Q000008:administration & dosage / Q000494:pharmacology; D023421:Models, Animal; D012212:Rheology / Q000187:drug effects; D013314:Stress, Mechanical; D034421:Sus scrofa; D023822:Tissue Engineering / Q000379:methods*; D014182:Transplantation, Autologous; D014682:Vena Cava, Inferior / Q000187:drug effects / Q000502:physiology
chemical_list: D001596:Benzylamines; D000080027:Cyclams; D006571:Heterocyclic Compounds; C088327:plerixafor
keywords: AMD3100; biocompatible materials; cell Isolation; cell therapy; endothelial cells; endothelial progenitor cells; swine; thrombosis; tissue engineering