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Array-Based Platform To Select, Release, and Capture Epstein-Barr Virus-Infected Cells Based on Intercellular

Peter J Attayek, Sally A Hunsucker1, Yuli Wang2

  • 1Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine , Chapel Hill, North Carolina 27599, United States.

Analytical Chemistry
|November 13, 2015
PubMed
Summary
This summary is machine-generated.

Microraft arrays enable cell separation based on complex phenotypes like weak adhesion, without needing known markers. This method efficiently isolates nonadherent single cells for culture and analysis.

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Microfluidics

Background:

  • Separating cells based on complex phenotypes, such as weak intercellular adhesion, is challenging.
  • Traditional cell separation methods often require knowledge of specific cell-surface markers or intracellular proteins.
  • Isolating nonadherent cells presents difficulties due to their inability to bind to culture surfaces.

Purpose of the Study:

  • To develop and validate a novel method for selecting and separating cells based on complex phenotypes, specifically weak intercellular adhesion.
  • To enable the isolation of nonadherent cells without requiring cell attachment to a surface.
  • To demonstrate the versatility of microraft arrays for cell separation and analysis.

Main Methods:

  • Development of microraft arrays for cell selection and separation.
  • A gelatin encapsulation method to immobilize and release nonadherent cells.
  • A semiautomated device for efficient microraft collection and cell release.
  • Isolation of Epstein-Barr virus-infected lymphoblastoid cell lines (EBV-LCL) based on intercellular adhesion.

Main Results:

  • 100% ± 0% microraft collection efficiency and a 5-fold increase in throughput using the semiautomated device.
  • 100% ± 0% isolation efficiency of non-surface-attached single cells with 96% ± 4% postsort single-cell cloning efficiency.
  • 87% ± 3% postsort viability for isolated EBV-LCL cells, with 100% ± 0% sorting efficiency.
  • Gene expression analysis showed no difference in EBNA-2 expression between adhesive and nonadhesive cells.

Conclusions:

  • Microraft arrays provide a versatile platform for separating cells based on complex and poorly understood phenotypes.
  • The gelatin encapsulation method effectively enables the isolation and culture of nonadherent cells.
  • This technology offers high efficiency and viability for cell isolation and downstream analysis, including genetic studies.