title: P-Selectin coated microtube for enrichment of CD34+ hematopoietic stem and progenitor cells from human bone marrow.
abstract: BACKGROUND
  Enrichment and purification of hematopoietic stem and progenitor cells (HSPCs) is important in transplantation therapies for hematologic disorders and in basic stem cell research. Primitive CD34+ HSPCs have demonstrated stronger rolling adhesion on selectins than mature CD34- mononuclear cells (MNCs). We have exploited this differential rolling behavior to capture and purify HSPCs from bone marrow by perfusing MNCs through selectin-coated microtubes.
  
  
  METHODS
  Bone marrow MNCs were perfused through the cell-capture microtubes coated with adhesion molecules. We washed the device lumen and visualized and estimated captured cells by video microscopy. Adherent cells were eluted by high shear, calcium-free buffer, and air embolism. We used immunofluorescence staining followed by flow cytometry to analyze CD34+ HSPCs.
  
  
  RESULTS
  CD34+ HSPC purity of cells captured in adhesion molecule-coated devices was significantly higher than the fraction of CD34+ cells found in bone marrow MNCs [mean (SE) 2.5% (0.8%)]. P-selectin-coated surfaces yielded 16% to 20% CD34+ cell purity, whereas antibody-coated surfaces yielded 12% to 18%. Although CD34+ cell purity was comparable between selectin and antibody surfaces, the total number of CD34+ HSPCs captured was significantly higher in P-selectin devices (approximately 5.7 x 10(4) to 7.1 x 10(4)) than antibody devices (approximately 1.74 x 10(4) to 2.61 x 10(4)).
  
  
  CONCLUSIONS
  P-selectin can be used in a compact flow device to capture HSPCs. Selectin-mediated capture of CD34+ HSPCs resulted in enrichment approximately 8-fold higher than the CD34+ cell population from bone marrow MNCs. This study supports the hypothesis that flow-based, adhesion molecule-mediated capture may be a viable alternative approach to the capture and purification of HSPCs.
authors.affiliation: Department of Biomedical Engineering, Wilmot Cancer Center, University of Rochester, Rochester, NY 14627, USA.
authors.forename: Srinivas D
authors.initials: SD
authors.lastname: Narasipura
authors.forename: Joel C
authors.initials: JC
authors.lastname: Wojciechowski
authors.forename: Nichola
authors.initials: N
authors.lastname: Charles
authors.forename: Jane L
authors.initials: JL
authors.lastname: Liesveld
authors.forename: Michael R
authors.initials: MR
authors.lastname: King
mesh_terms: D000911:Antibodies, Monoclonal; D018952:Antigens, CD34 / Q000276:immunology / Q000378:metabolism*; D002448:Cell Adhesion; D002465:Cell Movement; D002469:Cell Separation / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism; D019007:P-Selectin / Q000235:genetics; D011993:Recombinant Fusion Proteins
chemical_list: D000911:Antibodies, Monoclonal; D018952:Antigens, CD34; D019007:P-Selectin; D011993:Recombinant Fusion Proteins