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Related Experiment Video

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Flotation-Based T Cell Isolation, Activation, and Expansion from Human Peripheral Blood Mononuclear Cell Samples Using Microbubbles
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Buoyancy-activated cell sorting using targeted biotinylated albumin microbubbles.

Yu-Ren Liou1, Yu-Hsin Wang1, Chia-Ying Lee1

  • 1Graduate Institute of Biomedical Electronics and Bioinformatics, National Taiwan University, Taipei, Taiwan.

Plos One
|May 21, 2015
PubMed
Summary
This summary is machine-generated.

This study introduces buoyancy-activated cell sorting using targeted biotinylated albumin microbubbles (biotin-MBs) for gentle cell isolation. This novel method effectively separates MDA-MB-231 breast cancer cells with high purity and recovery rates.

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Biomedical Engineering

Background:

  • Conventional cell isolation methods like fluorescence-activated and magnetic-activated cell sorting risk cell damage due to mechanical forces.
  • There is a need for non-destructive cell isolation techniques for applications in research and clinical trials.
  • Albumin microbubbles (MBs) are biocompatible ultrasound contrast agents suitable for modification.

Purpose of the Study:

  • To develop and evaluate a novel, gentle cell isolation method: buoyancy-activated cell sorting (BACS).
  • To assess the efficacy of targeted biotinylated albumin microbubbles (biotin-MBs) conjugated with anti-CD44 antibodies for separating MDA-MB-231 breast cancer cells.
  • To compare the performance of biotin-MBs with conventional avidin-incorporated albumin MBs (avidin-MBs).

Main Methods:

  • Preparation of biotinylated albumin microbubbles (biotin-MBs) with a mean diameter of 2 μm.
  • Conjugation of anti-CD44 antibodies to biotin-MBs via an avidin-biotin system to create targeted biotin-MBs.
  • Incubation of MDA-MB-231 cells with targeted biotin-MBs, followed by centrifugation and a 1-hour incubation at 4°C for separation.

Main Results:

  • Targeted biotin-MBs successfully separated MDA-MB-231 breast cancer cells, with over 90% collected in the MB layer at a MBs-to-cells ratio > 70:1.
  • Separation efficiency of targeted biotin-MBs was superior to that of targeted avidin-MBs.
  • High sorting purity (>84%) and recovery rate (up to 88%) were achieved for a heterogeneous cell population including CD44(+) MDA-MB-231 and CD44(-) MDA-MB-453 cells.

Conclusions:

  • Buoyancy-activated cell sorting using targeted biotin-MBs offers a highly efficient and gentle method for cell isolation.
  • This technique demonstrates significant potential for isolating cancer stem cells (CSCs), identified by CD44 expression, from tumor tissues.
  • Targeted biotin-MBs represent a promising tool for preclinical research and clinical applications requiring pure cell populations.