Biomedical Microdevices

, Volume 13, Issue 3, pp 415–430 | Cite as

Microfluidic cell culture chip with multiplexed medium delivery and efficient cell/scaffold loading mechanisms for high-throughput perfusion 3-dimensional cell culture-based assays

  • Song-Bin Huang
  • Min-Hsien Wu
  • Shih-Siou Wang
  • Gwo-Bin Lee
Article

Abstract

This study reports a microfluidic cell culture chip consisting of 48 microbioreactors for high-throughput perfusion 3-dimensional (3-D) cell culture-based assays. Its advantages include the capability for multiplexed and backflow-free medium delivery, and both efficient and high-throughput micro-scale, 3-D cell culture construct loading. In this work, the microfluidic cell culture chip is fabricated using two major processes, specifically, a computer-numerical-controlled (CNC) mold machining process and a polydimethylsiloxane (PDMS) replication process. The chip is composed of micropumps, microbioreactors, connecting microchannels and a cell/agarose scaffold loading mechanism. The performance of the new pneumatic micropumps and the cell/agarose scaffold loading mechanism has been experimentally evaluated. The experimental results show that this proposed multiplexed medium-pumping design is able to provide a uniform pumping rate ranging from 1.5 to 298.3 μl hr−1 without any fluid backflow and the resultant medium contamination. In addition, the simple cell/agarose loading method has been proven to be able to load the 3-D cell culture construct uniformly and efficiently in all 48 microbioreactors investigated. Furthermore, a micro-scale, perfusion, 3-D cell culture-based assay has been successfully demonstrated using this proposed cell culture chip. The experimental results are also compared to a similar evaluation using a conventional static 3-D cell culture with a larger scale culture. It is concluded that the choice of a cell culture format can influence assay results. As a whole, because of the inherent advantages of a miniaturized perfusion 3-D cell culture assay, the cell culture chip not only can provide a stable, well-defined and more biologically-meaningful culture environment, but it also features a low consumption of research resources. Moreover, due to the integrated medium pumping mechanism and the simple cell/agarose loading method, this chip is economical and time efficient. All of these traits are particularly useful for high-precision and high-throughput 3-D cell culture-based assays.

Keywords

Microfluidics Micropumps Microbioreactors Perfusion cell culture 3-D cell culture 

Supplementary material

10544_2011_9510_MOESM1_ESM.mpg (617 kb)
ESM 1(MPG 617 kb)
10544_2011_9510_MOESM2_ESM.mpg (3.6 mb)
ESM 2(MPG 3701 kb)

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Song-Bin Huang
    • 1
  • Min-Hsien Wu
    • 2
  • Shih-Siou Wang
    • 2
  • Gwo-Bin Lee
    • 3
  1. 1.Department of Engineering ScienceNational Cheng Kung UniversityTainanTaiwan
  2. 2.Graduate Institute of Biochemical and Biomedical EngineeringChang Gung UniversityTaoyuanTaiwan
  3. 3.Department of Power Mechanical EngineeringNational Tsing Hua UniversityHsinchuTaiwan

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