title: Patient-Derived Airway Secretion Dissociation Technique To Isolate and Concentrate Immune Cells Using Closed-Loop Inertial Microfluidics.
abstract: Assessment of airway secretion cells, both for research and clinical purposes, is a highly desired goal in patients with acute and chronic pulmonary diseases. However, lack of proper cell isolation and enrichment techniques hinder downstream evaluation and characterization of cells found in airway secretions. Here, we demonstrate a novel enrichment method to capture immune-related cells from clinical airway secretions using closed-loop separation of spiral inertial microfluidics (C-sep). By recirculating the output focusing stream back to the input reservoir and running continuously with a high flow processing rate, one can achieve optimal concentration, recovery and purity of airway immune cells from a large volume of diluent, which was not readily possible in the single-pass operation. Our method reproducibly recovers 94.0% of polymorphonuclear leukocytes (PMNs), with up to 105PMNs in clear diluted buffer from 50 μL of airway secretions obtained from mechanically ventilated patients. We show that C-sep isolated PMNs show higher neutrophil elastase (NE) release following activation by phorbol 12-myristate 13-acetate (PMA) than cells isolated by conventional mucolytic method. By capturing cells without chemically disrupting their potential function, our method is expected to expand the possibility of clinical in vitro cell based biological assays for various pulmonary diseases such as acute respiratory distress syndrome, pneumonia, cystic fibrosis, and bronchiectasis.
authors.affiliation: Research Laboratory of Electronics, ‡Department of Electrical Engineering and Computer Science, §Department of Biological Engineering, Massachusetts Institute of Technology , 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.|Department of Medicine and the ⊥Vascular Medicine Institute, University of Pittsburgh , NW628 Montefiore University Hospital, 3459 Fifth Avenue, Pittsburgh, Pennsylvania 15213, United States.
authors.forename: Hyunryul
authors.identifier: 0000-0003-3645-1930
authors.initials: H
authors.lastname: Ryu
authors.affiliation: Research Laboratory of Electronics, ‡Department of Electrical Engineering and Computer Science, §Department of Biological Engineering, Massachusetts Institute of Technology , 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.|Department of Medicine and the ⊥Vascular Medicine Institute, University of Pittsburgh , NW628 Montefiore University Hospital, 3459 Fifth Avenue, Pittsburgh, Pennsylvania 15213, United States.
authors.forename: Kyungyong
authors.initials: K
authors.lastname: Choi
authors.affiliation: Research Laboratory of Electronics, ‡Department of Electrical Engineering and Computer Science, §Department of Biological Engineering, Massachusetts Institute of Technology , 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.|Department of Medicine and the ⊥Vascular Medicine Institute, University of Pittsburgh , NW628 Montefiore University Hospital, 3459 Fifth Avenue, Pittsburgh, Pennsylvania 15213, United States.
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Qu
authors.affiliation: Research Laboratory of Electronics, ‡Department of Electrical Engineering and Computer Science, §Department of Biological Engineering, Massachusetts Institute of Technology , 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.|Department of Medicine and the ⊥Vascular Medicine Institute, University of Pittsburgh , NW628 Montefiore University Hospital, 3459 Fifth Avenue, Pittsburgh, Pennsylvania 15213, United States.
authors.forename: Taehong
authors.identifier: 0000-0002-6635-2557
authors.initials: T
authors.lastname: Kwon
authors.affiliation: Research Laboratory of Electronics, ‡Department of Electrical Engineering and Computer Science, §Department of Biological Engineering, Massachusetts Institute of Technology , 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.|Department of Medicine and the ⊥Vascular Medicine Institute, University of Pittsburgh , NW628 Montefiore University Hospital, 3459 Fifth Avenue, Pittsburgh, Pennsylvania 15213, United States.
authors.forename: Janet S
authors.initials: JS
authors.lastname: Lee
authors.affiliation: Research Laboratory of Electronics, ‡Department of Electrical Engineering and Computer Science, §Department of Biological Engineering, Massachusetts Institute of Technology , 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.|Department of Medicine and the ⊥Vascular Medicine Institute, University of Pittsburgh , NW628 Montefiore University Hospital, 3459 Fifth Avenue, Pittsburgh, Pennsylvania 15213, United States.
authors.forename: Jongyoon
authors.initials: J
authors.lastname: Han
mesh_terms: D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D004229:Dithiothreitol / Q000494:pharmacology; D006801:Humans; D019272:Leukocyte Elastase / Q000378:metabolism; D008171:Lung Diseases / Q000276:immunology / Q000473:pathology; D044085:Microfluidics; D009077:Mucins / Q000378:metabolism; D009504:Neutrophils / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D013183:Sputum / Q000166:cytology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D009077:Mucins; D019272:Leukocyte Elastase; D013755:Tetradecanoylphorbol Acetate; D004229:Dithiothreitol