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Patient-Derived Airway Secretion Dissociation Technique To Isolate and Concentrate Immune Cells Using Closed-Loop
Hyunryul Ryu1,2, Kyungyong Choi1,2, Yanyan Qu1,2
1Research 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.
Insights
A novel microfluidic method effectively isolates immune cells from airway secretions, improving analysis for pulmonary diseases. This technique enhances cell purity and function for better research and clinical applications.
Area of Science:
- Biomedical Engineering
- Cell Biology
- Pulmonary Medicine
Background:
- Assessing airway secretion cells is crucial for diagnosing and treating pulmonary diseases.
- Current cell isolation methods are insufficient for downstream analysis, limiting research and clinical applications.
Purpose of the Study:
- To develop and validate a novel closed-loop microfluidic enrichment method for airway immune cells.
- To improve the purity, recovery, and functional integrity of cells from clinical airway secretions.
Main Methods:
- Utilized closed-loop separation of spiral inertial microfluidics (C-sep) for continuous cell enrichment.
- Processed clinical airway secretions from mechanically ventilated patients.
- Compared C-sep isolated polymorphonuclear leukocytes (PMNs) with those from conventional methods.
Main Results:
- Achieved reproducible recovery of 94.0% of PMNs with high purity.
- Isolated up to 10^5 PMNs from 50 μL of airway secretions.
- C-sep isolated PMNs exhibited enhanced neutrophil elastase release post-activation compared to conventional methods.
Conclusions:
- C-sep offers an efficient and functional method for isolating airway immune cells from clinical samples.
- This technique supports advanced in vitro cell-based assays for pulmonary diseases.
- C-sep has the potential to improve diagnostics and therapeutic strategies for conditions like ARDS, pneumonia, and cystic fibrosis.
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 105 PMNs 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.
