title: Bone matrix vesicle-bound alkaline phosphatase for the assessment of peripheral blood admixture to human bone marrow aspirates.
abstract: OBJECTIVE
  Peripheral blood (PB) admixture should be minimized during numerical and functional, as well as cytokinetic analysis of bone marrow (BM) aspirates for research purposes. Therefore, purity assessment of the BM aspirate should be performed in advance. We investigated whether bone matrix vesicle (BMV)-bound bone alkaline phosphatase (ALP) could serve as a marker for the purity of BM aspirates.
  
  
  RESULTS
  Total ALP activity was significantly higher in BM serum (97 (176-124)U/L, median (range)) compared to PB serum (63 (52-73)U/L, p < 0.001). Agarose gel electrophoresis showed a unique bone ALP fraction in BM, which was absent in PB. Native polyacrylamide gel electrophoresis revealed the high molecular weight of this fraction, corresponding with membrane-bound ALP from bone matrix vesicles (BMV), as evidenced by electron microscopy. A serial PB admixture experiment of bone cylinder supernatant samples, rich in BMV-bound ALP, confirmed the sensitivity of this proposed quality assessment method. Furthermore, a BMV ALP fraction of ≥ 15% is suggested as cut-off value for minimal BM quality. Moreover, the BM purity declines rapidly with larger aspirated BM volumes.
  
  
  CONCLUSIONS
  The exclusive presence of BMV-bound ALP in BM could serve as a novel marker to assess purity of BM aspirates.
authors.affiliation: Laboratory of Cellular and Molecular Cardiology, Department of Cardiology, Antwerp University Hospital, Antwerp, Belgium; Cardiovascular Diseases, Department of Translational Pathophysiological Research, University of Antwerp, Antwerp, Belgium. Electronic address: Evelien.Nollet@uantwerpen.be.
authors.forename: Evelien
authors.initials: E
authors.lastname: Nollet
authors.affiliation: Laboratory of Cellular and Molecular Cardiology, Department of Cardiology, Antwerp University Hospital, Antwerp, Belgium; Cardiovascular Diseases, Department of Translational Pathophysiological Research, University of Antwerp, Antwerp, Belgium; Department of Cardiology, Antwerp University Hospital, Antwerp, Belgium.
authors.forename: Emeline M
authors.initials: EM
authors.lastname: Van Craenenbroeck
authors.affiliation: Laboratory of Physiopharmacology, Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, Belgium.
authors.forename: Wim
authors.initials: W
authors.lastname: Martinet
authors.affiliation: Department of Cardiac Surgery, Antwerp University Hospital, Antwerp, Belgium.
authors.forename: Inez
authors.initials: I
authors.lastname: Rodrigus
authors.affiliation: Department of Cardiac Surgery, Antwerp University Hospital, Antwerp, Belgium.
authors.forename: Dina
authors.initials: D
authors.lastname: De Bock
authors.affiliation: Vaccine & Infectious Disease Institute, Laboratory of Experimental Hematology, University of Antwerp, Antwerp, Belgium.
authors.forename: Zwi
authors.initials: Z
authors.lastname: Berneman
authors.affiliation: Laboratory of Cell Biology and Histology, University of Antwerp, Antwerp, Belgium.
authors.forename: Isabel
authors.initials: I
authors.lastname: Pintelon
authors.affiliation: Department of Hepatobiliary, Transplantation and Endocrine Surgery, Antwerp University Hospital, Antwerp, Belgium; Laboratory of Experimental Surgery, University of Antwerp, Antwerp, Belgium.
authors.forename: Dirk
authors.initials: D
authors.lastname: Ysebaert
authors.affiliation: Laboratory of Cellular and Molecular Cardiology, Department of Cardiology, Antwerp University Hospital, Antwerp, Belgium; Cardiovascular Diseases, Department of Translational Pathophysiological Research, University of Antwerp, Antwerp, Belgium; Department of Cardiology, Antwerp University Hospital, Antwerp, Belgium.
authors.forename: Christiaan J
authors.initials: CJ
authors.lastname: Vrints
authors.affiliation: Laboratory of Cellular and Molecular Cardiology, Department of Cardiology, Antwerp University Hospital, Antwerp, Belgium; Cardiovascular Diseases, Department of Translational Pathophysiological Research, University of Antwerp, Antwerp, Belgium; Department of Cardiology, Antwerp University Hospital, Antwerp, Belgium.
authors.forename: Viviane M
authors.initials: VM
authors.lastname: Conraads
authors.affiliation: Department of Clinical Chemistry, Antwerp University Hospital, Antwerp, Belgium; Biochemistry, Department of Translational Pathophysiological Research, University of Antwerp, Antwerp, Belgium.
authors.forename: Viviane O M
authors.initials: VO
authors.lastname: Van Hoof
mesh_terms: D000368:Aged; D000469:Alkaline Phosphatase / Q000032:analysis* / Q000145:classification / Q000378:metabolism; D001707:Biopsy, Needle / Q000592:standards*; D001853:Bone Marrow / Q000502:physiology* / Q000648:ultrastructure; D016026:Bone Marrow Transplantation; D001857:Bone Matrix / Q000201:enzymology* / Q000648:ultrastructure; D006348:Cardiac Surgical Procedures; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000201:enzymology; D008297:Male; D008854:Microscopy, Electron; D008875:Middle Aged; D011485:Protein Binding; D011786:Quality Control; D014182:Transplantation, Autologous
chemical_list: D000469:Alkaline Phosphatase
keywords: Alkaline phosphatase; Blood; Bone marrow; Bone marrow examination/methods; Quality control