title: Tag-free microfluidic separation of cells against multiple markers.
abstract: Conventional cell separation against multiple markers generally requires the attachment of antibody tags, typically fluorescent or magnetic, to selected cell types in a heterogeneous suspension. This work describes how such separation can be accomplished in a series of microfluidic systems without the need for such tags. Two capture stages containing antibody-functionalized alginate hydrogels are utilized for the isolation of CD34+ and Flk1+ cells from untreated, whole human blood. The capture-release capability of these degradable coatings is harnessed by a mixing chamber and a simple valving system such that the suspension emerging from the first capture stage is prepared for the second capture stage for further enrichment. With this configuration, we demonstrate the isolation of CD34+/Flk1+ endothelial progenitor cells from blood enabled by the depletion of CD34+/Flk1-hematopoietic stem cells population. This ability to achieve isolation of cells against multiple markers in an untagged separation method is of particular significance in applications involving cell implantation-based therapeutics including tissue engineering and molecular analysis.
authors.affiliation: Department of Chemical Engineering, Northeastern University, Boston, Massachusetts 02115, United States.
authors.forename: Adam
authors.initials: A
authors.lastname: Hatch
authors.forename: Danielle M
authors.initials: DM
authors.lastname: Pesko
authors.forename: Shashi K
authors.initials: SK
authors.lastname: Murthy
mesh_terms: D000464:Alginates / Q000737:chemistry; D018952:Antigens, CD34 / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002469:Cell Separation; D005434:Flow Cytometry; D020723:Glucuronic Acid / Q000737:chemistry; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006603:Hexuronic Acids / Q000737:chemistry; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D046210:Microfluidic Analytical Techniques; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism
chemical_list: D000464:Alginates; D018952:Antigens, CD34; D015415:Biomarkers; D006603:Hexuronic Acids; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D020723:Glucuronic Acid; D040301:Vascular Endothelial Growth Factor Receptor-2