title: Perfusion method for harvesting bone marrow cells from dogs.
abstract: OBJECTIVE
  To compare composition and colony formation of bone marrow mononuclear cells (BMMCs) harvested from dogs by means of a new perfusion method and the conventional aspiration method.
  
  
  METHODS
  7 healthy adult Beagles.
  
  
  METHODS
  BMMCs were collected from the humeri and femurs of Beagles via perfusion and aspiration methods. Flow cytometric analysis was performed to quantify the presence of contaminant cells from the peripheral blood and the percentage of CD34+ progenitor cells in the BMMCs. A CFU assay was conducted to determine the number of progenitor cells in the BMMCs.
  
  
  RESULTS
  The perfusion method was safely performed in all 7 dogs. Flow cytometric analysis revealed that the percentages of contaminant CD3+CD4+, CD3+CD8+, and CD21 + lymphocytes in BMMCs obtained via perfusion were significantly lower than percentages obtained via aspiration. The percentage of CD34+ cells obtained via perfusion was significantly higher than that obtained via aspiration. In addition, perfusion yielded a significantly higher CFU count than did aspiration.
  
  
  CONCLUSIONS
  The perfusion method used in this study can minimize the contamination of bone marrow samples with peripheral blood and was a more efficient means for collecting canine bone marrow progenitor cells than the conventional aspiration method. Therefore, the perfusion method can be more suitable than aspiration for harvesting bone marrow cells for transplantation in dogs.
authors.affiliation: Department of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, University of Tokyo, 1-1-1, Yayoi, Bunkyo-ku, Tokyo, Japan.
authors.forename: Masahiko
authors.initials: M
authors.lastname: Sato
authors.forename: Yuko
authors.initials: Y
authors.lastname: Goto-Koshino
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Mochizuki
authors.forename: Yasuhito
authors.initials: Y
authors.lastname: Fujino
authors.forename: Koichi
authors.initials: K
authors.lastname: Ohno
authors.forename: Hajime
authors.initials: H
authors.lastname: Tsujimoto
mesh_terms: D000818:Animals; D018952:Antigens, CD34; D001707:Biopsy, Needle / Q000379:methods / Q000662:veterinary; D001854:Bone Marrow Cells / Q000166:cytology*; D002469:Cell Separation / Q000379:methods / Q000662:veterinary*; D004285:Dogs; D005434:Flow Cytometry; D010477:Perfusion / Q000379:methods / Q000662:veterinary*; D013234:Stem Cells
chemical_list: D018952:Antigens, CD34