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Unsupervised capture and profiling of rare immune cells using multi-directional magnetic ratcheting.

Abstract
Immunotherapies (IT) require induction, expansion, and maintenance of specific changes to a patient's immune cell repertoire which yield a therapeutic benefit. Recently, mechanistic understanding of these changes at the cellular level has revealed that IT results in complex phenotypic transitions in target cells, and that therapeutic effectiveness may be predicted by monitoring these transitions during therapy. However, monitoring will require unique tools that enable capture, manipulation, and profiling of rare immune cell populations. In this study, we introduce a method of automated and unsupervised separation and processing of rare immune cells, using high-force and multidimensional magnetic ratcheting (MR). We demonstrate capture of target immune cells using samples with up to 1 : 10 000 target cell to background cell ratios from input volumes as small as 25 microliters (i.e. a low volume and low cell frequency sample sparing assay interface). Cell capture is shown to achieve up to 90% capture efficiency and purity, and captured cell analysis is shown using both on-chip culture/activity assays and off-chip ejection and nucleic acid analysis. These results demonstrate that multi-directional magnetic ratcheting offers a unique separation system for dealing with blood cell samples that contain either rare cells or significantly small volumes, and the "sample sparing" capability leads to an expanded spectrum of parameters that can be measured. These tools will be paramount to advancing techniques for immune monitoring under conditions in which both the sample volume and number of antigen-specific target cells are often exceedingly small, including during IT and treatment of allergy, asthma, autoimmunity, immunodeficiency, cell based therapy, transplantation, and infection.
AuthorsColeman Murray, Hiromi Miwa, Manjima Dhar, Da Eun Park, Edward Pao, Jessica Martinez, Sireesha Kaanumale, Evelina Loghin, John Graf, Khadir Raddassi, William W Kwok, David Hafler, Chris Puleo, Dino Di Carlo
JournalLab on a chip (Lab Chip) Vol. 18 Issue 16 Pg. 2396-2409 (08 07 2018) ISSN: 1473-0189 [Electronic] England
PMID30039125 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
Chemical References
  • Cytokines
Topics
  • Cell Separation (instrumentation)
  • Cytokines (metabolism)
  • Humans
  • Immune System (cytology)
  • Magnetic Fields

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