An automatic microfluidic system for rapid screening of cancer stem-like cell-specific aptamers

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• 作者：Chen-Hsun Weng (1)
  I-Shan Hsieh (2)
  Lien-Yu Hung (5)
  Hsin-I Lin (4)
  Shu-Chu Shiesh (4)
  Yuh-Ling Chen (3)
  Gwo-Bin Lee (5)
  
• 关键词：Cell ; SELEX ; Microfluidic ; Magnetic bead ; Aptamer ; Cancer stem cell
• 刊名：Microfluidics and Nanofluidics
• 出版年：2013
• 出版时间：4 - March 2013
• 年：2013
• 卷：14
• 期：3
• 页码：753-765
• 全文大小：930KB
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• 作者单位：Chen-Hsun Weng (1)
  I-Shan Hsieh (2)
  Lien-Yu Hung (5)
  Hsin-I Lin (4)
  Shu-Chu Shiesh (4)
  Yuh-Ling Chen (3)
  Gwo-Bin Lee (5)
  
  1. Department of Microbiology and Immunology, National Cheng Kung University, Tainan, 70101, Taiwan
  2. Institute of Basic Medical Sciences, National Cheng Kung University, Tainan, 70101, Taiwan
  5. Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, 30013, Taiwan
  4. Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University, Tainan, 70101, Taiwan
  3. Institute of Oral Medicine, National Cheng Kung University, Tainan, 70101, Taiwan
  
• ISSN：1613-4990

文摘

This study reports a microfluidic system which automatically performs the systematic evolution of ligands by exponential enrichment (SELEX) process for rapid screening of aptamers which are specific to cancer stem-like cells. The system utilizes magnetic bead-based techniques to select DNA aptamers and has several advantages including a rapid, automated screening process, and less consumption of cells and reagents. By integrating a microfluidic control module, a magnetic bead-based aptamer extraction module, and a temperature control module, the entire Cell-SELEX process can be performed in a shorter period of time. Compared with the traditional Cell-SELEX process, this microfluidic system is more efficient and consumes fewer sample volumes. It only takes approximately 3?days for an entire Cell-SELEX process with 15 screening runs, which is relatively faster than that of a traditional Cell-SELEX process (1?week for 15 rounds). The binding affinity of this resulting specific aptamer was measured by a flow cytometric analysis to have a dissociation constant (K d) of 15.32?nM. The capture rate for cancer stem-like cells using the specific aptamer-conjugated bead is better than that using Ber-EP4 antibody-conjugated bead. This microfluidic system may provide a powerful platform for the rapid screening of cell-specific aptamers.