High-Throughput, low-loss, low-cost, and label-free cell separation using electrophysiology-Activated cell enrichment

Shabnam A. Faraghat, Kai F. Hoettges, Max K. Steinbach, Daan R. Van Der Veen, William J. Brackenbury, Erin A. Henslee, Fatima H. Labeed, Michael P. Hughes

Research output: Contribution to journalArticlepeer-review

85 Citations (Scopus)

Abstract

Currently, cell separation occurs almost exclusively by density gradient methods and by fluorescence-And magnetic-Activated cell sorting (FACS/MACS). These variously suffer from lack of specificity, high cell loss, use of labels, and high capital/operating cost. We present a dielectrophoresis (DEP)-based cell-separation method, using 3D electrodes on a low-cost disposable chip; one cell type is allowed to pass through the chip whereas the other is retained and subsequently recovered. The method advances usability and throughput of DEP separation by orders of magnitude in throughput, efficiency, purity, recovery (cells arriving in the correct output fraction), cell losses (those which are unaccounted for at the end of the separation), and cost. The system was evaluated using three example separations: live and dead yeast; human cancer cells/red blood cells; and rodent fibroblasts/red blood cells. A single-pass protocol can enrich cells with cell recovery of up to 91.3% at over 300,000 cells per second with >3% cell loss. A twopass protocol can process 300,000,000 cells in under 30 min, with cell recovery of up to 96.4% and cell losses below 5%, an effective processing rate >160,000 cells per second. A three-step protocol is shown to be effective for removal of 99.1% of RBCs spiked with 1%cancer cellswhilemaintaining a processing rate of ∼170,000 cells per second. Furthermore, the self-contained and low-cost nature of the separator device means that it has potential application in lowcontamination applications such as cell therapies, where good manufacturing practice compatibility is of paramount importance.

Original languageEnglish
Pages (from-to)4591-4596
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume114
Issue number18
DOIs
Publication statusPublished - May 2 2017
Externally publishedYes

Keywords

  • DEP
  • Dielectrophoresis
  • FACS
  • Lab on a chip
  • MACS

ASJC Scopus subject areas

  • General

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