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Presented by: Casey Hua, Danyi Wu

Isolating highly enriched populations of circulating epithelial cells and other rare cells from blood using a magnetic sweeper device.

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Presented by: Casey Hua, Danyi Wu

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  1. Isolating highly enriched populations of circulating epithelial cells and other rare cells from blood using a magnetic sweeper device AmirAli H. Talasaz, Ashley A. Powell, David E. Huber, James G. Berbee, Kyung-Ho Roh, Wong Yu, Wenzhong Xiao, Mark M. Davis, R. Fabian Pease, Michael N. Mindrinos, Stefanie S. Jeffrey, and Ronald W. Davis Presented by: Casey Hua, Danyi Wu

  2. Motivation • Molecular Profiling Applications • Improvement of: • Cancer prognosis/treatment  rare circulating epithelial cells (CEpCs) • Immune monitoring  antigen-specific lymphocytes • Prenatal diagnosis  fetal cells

  3. Motivation Problem: Current CEpC isolation platforms lack purification efficiency • Solution: MagSweeper • High capture rate • High purity • Enriches target cells (108x)

  4. MagSweeper Process • Pre-label diluted blood samples with magnetic particles 2. 3. 4. 5. Repeat to eliminate background cells  improves purity at some cost to capture efficiency

  5. MagSweeper Simulation Magnetic flux density Magnetic force-driven cell velocity field Cell capture boundary

  6. MagSweeper Simulation cont’d Fluid velocity/motion Cell trapping profiles Gap of uncaptured particles due to fluid motion pushing particles ahead of magnet

  7. MagSweeper Performance • Enrichment Studies • Rare Immunological Model Cells from a Mixed Sample (HLA-A2) • Capture Efficiency • Purity • MCF7 Cancer Cells Spiked in Blood Samples • Performance: isolating Circulating Epithelial Cells (CEpCs) • CEpCs from Breast Cancer Patients • Application: CEpC isolation from Epithelial Cancers patients • Perturbation of Cellular Gene Expression

  8. (1) Rare HLA-A2+ cells in Mixed Samples Enrichment by ~2.5x105

  9. (2) MCF7 Cancer Cells Spiked in Blood Samples • Combining MagSweeper enrichment with visual inspection and extraction of cells based on size: • increase cell purification of the MCF7 cells from 51% to 100% purity • similar cell capture rates from 62%± 7% of 59%± 27% MCF7 cells isolated by MagSweeper with brightfield and fluorescence imaging.

  10. (3) CEpCs in Breast Cancer Patients • Found CEpCs in 47 of 47 samples from patients with metastatic disease • 12±23 CEpCs per 9mL blood sample

  11. MagSweeper isolation protocol does not perturb gene expression minimal changes in cell profiles: 94% of probe sets have changes <50% Fig. S2. Fold changes in logarithmic scale between the gene expression levels of cultured cells before and after MagSweeper protocol.

  12. Advantages of MagSweeper • High enrichment • No prior processing of blood required • Flexibility in starting sample volume & process scalability • Further Optimization: • Extension of the sweep to capture remaining particles in gap left by fluid motion • Refine magnet shape to yield more uniform magnetic field gradient

  13. Significance • Diagnose malignancy from blood • Improved cancer prognosis: CEpC numbers correlate better with cancer progression than imaging, lymph node biopsy, and serum biomarker analysis • Enrich CEpCs for molecular analysis • Identify surrogate endpoints for evaluation in clinical trials • Extract individual CEpCs • characterization of single cells for subpopulation studies on heterogenous CEpCs

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