The separation of peripheral blood mononuclear cells (PBMCs) into constituent blood cell types is a vital step to obtain immune cells for autologous cell therapies. The ability to separate PBMCs using label-free microfluidic techniques, based on differences in biomechanical properties, can have a number of benefits over other conventional techniques, including lower cost, ease of use, and avoidance of animal-derived labeling antibodies. Here, we report a microfluidic device that uses compressive diagonal ridges to separate PBMCs into highly pure samples of viable and functional lymphocytes. The technique utilizes the differences in the biophysical properties of PBMC sub-populations to direct the lymphocytes and monocytes into separate outlets. The biophysical properties of the monocytes and lymphocytes from healthy donors were first characterized using atomic force microscopy. Lymphocytes were found to be significantly stiffer than monocytes, with a mean cell stiffness of 1495 and 931 Pa, respectively. The differences in biophysical properties resulted in distinct trajectories through the microchannel terminating at different outlets, resulting in a lymphocyte sample with purity and viability both greater than 96% with no effect on the cells’ ability to produce interferon gamma, a cytokine crucial for innate and adaptive immunity.
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September 2023
Research Article|
September 18 2023
Label-free microfluidic isolation of functional and viable lymphocytes from peripheral blood mononuclear cells
Special Collection:
Microfluidics and Nanofluidics for Immunotherapy
Abhishek Raj
;
Abhishek Raj
a)
(Conceptualization, Data curation, Investigation, Methodology, Project administration, Validation, Visualization, Writing – original draft)
1
George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology
, 801 Ferst Drive, Atlanta, Georgia 30332-0405, USA
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Katily Ramirez
;
Katily Ramirez
(Data curation, Formal analysis, Investigation, Writing – original draft, Writing – review & editing)
2
School of Chemistry & Biochemistry, Georgia Institute of Technology
, 901 Atlantic Drive, Atlanta, Georgia 30332-0400, USA
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Katherine M. Young
;
Katherine M. Young
(Data curation, Formal analysis, Methodology, Validation, Writing – original draft)
3
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology
, 313 Ferst Drive, Atlanta, Georgia 30332-0535, USA
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Nicholas Stone
;
Nicholas Stone
(Data curation, Formal analysis, Software)
1
George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology
, 801 Ferst Drive, Atlanta, Georgia 30332-0405, USA
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Peter Shankles
;
Peter Shankles
(Formal analysis, Methodology)
1
George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology
, 801 Ferst Drive, Atlanta, Georgia 30332-0405, USA
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Mehdia Nadeem Rajab Ali
;
Mehdia Nadeem Rajab Ali
(Project administration, Visualization, Writing – review & editing)
3
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology
, 313 Ferst Drive, Atlanta, Georgia 30332-0535, USA
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Anthony Malik Compton
;
Anthony Malik Compton
(Project administration, Visualization, Writing – review & editing)
3
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology
, 313 Ferst Drive, Atlanta, Georgia 30332-0535, USA
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Wilbur Lam
;
Wilbur Lam
(Resources)
3
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology
, 313 Ferst Drive, Atlanta, Georgia 30332-0535, USA
4
Winship Cancer Institute, Emory School of Medicine
, 1365 Clifton NE Rd., Atlanta, Georgia 30322, USA
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Alexander Alexeev
;
Alexander Alexeev
(Conceptualization, Supervision)
1
George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology
, 801 Ferst Drive, Atlanta, Georgia 30332-0405, USA
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Todd Sulchek
Todd Sulchek
b)
(Conceptualization, Funding acquisition, Project administration, Resources, Supervision)
1
George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology
, 801 Ferst Drive, Atlanta, Georgia 30332-0405, USA
2
School of Chemistry & Biochemistry, Georgia Institute of Technology
, 901 Atlantic Drive, Atlanta, Georgia 30332-0400, USA
b)Author to whom correspondence should be addressed: todd.sulchek@me.gatech.edu
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b)Author to whom correspondence should be addressed: todd.sulchek@me.gatech.edu
a)
Present address: Department of Mechanical Engineering, Indian Institute of Technology Patna, Bihta, Bihar 801106, India.
Biomicrofluidics 17, 054102 (2023)
Article history
Received:
June 07 2023
Accepted:
August 28 2023
Citation
Abhishek Raj, Katily Ramirez, Katherine M. Young, Nicholas Stone, Peter Shankles, Mehdia Nadeem Rajab Ali, Anthony Malik Compton, Wilbur Lam, Alexander Alexeev, Todd Sulchek; Label-free microfluidic isolation of functional and viable lymphocytes from peripheral blood mononuclear cells. Biomicrofluidics 1 September 2023; 17 (5): 054102. https://doi.org/10.1063/5.0161047
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