Cell separation mediated by differential rolling adhesion.
Recently, we showed a correlation between the maturity of hematopoietic stem and progenitor cells during development and rolling efficiency on selectins. These findings motivated us to explore a novel separation that exploits differences in selectin-mediated rolling adhesion between populations of cells. We extend the use of a previously developed cell-free system to study the separation of populations of sialyl Lewis x (sLe(x))-coated microspheres designed to roll with different average velocities on L-selectin chimeric substrates under well-defined flow. Results show that a separation that exploits differences in average rolling velocities between cell or microsphere populations is attainable. Excellent recovery and purity values for the slower rolling, or more desirable, populations are obtained and can be estimated from rolling velocity measurements. We also assess the feasibility of a selectin-mediated separation of adult bone marrow cell populations using previously obtained rolling velocity and rolling flux data for CD34+ and CD34- adult bone marrow cells on L-selectin substrates. We believe that a cell separation mediated by differential rolling adhesion can be used to enrich populations of hematopoietic stem and progenitor cells from an adult bone marrow cell preparation and that this method possesses several major advantages over existing antibody-mediated cell-affinity chromatography technologies.
Department of Bioengineering and Institute for Medicine and Engineering, University of Pennsylvania, 3320 Smith Walk, Philadelphia, Pennsylvania 19104, USA.
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Greenberg
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Hammer
D000328:Adult; D018952:Antigens, CD34 / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000502:physiology; D002448:Cell Adhesion; D002469:Cell Separation / Q000379:methods*; D004867:Equipment Design; D005240:Feasibility Studies; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000502:physiology; D006801:Humans; D019041:L-Selectin / Q000378:metabolism; D008863:Microspheres; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D018952:Antigens, CD34; D011993:Recombinant Fusion Proteins; D019041:L-Selectin