title: Microfluidic cell culture chip with multiplexed medium delivery and efficient cell/scaffold loading mechanisms for high-throughput perfusion 3-dimensional cell culture-based assays.
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.
authors.affiliation: Department of Engineering Science, National Cheng Kung University, Tainan, Taiwan.
authors.forename: Song-Bin
authors.initials: SB
authors.lastname: Huang
authors.forename: Min-Hsien
authors.initials: MH
authors.lastname: Wu
authors.forename: Shih-Siou
authors.initials: SS
authors.lastname: Wang
authors.forename: Gwo-Bin
authors.initials: GB
authors.lastname: Lee
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation*; D019902:Chondrocytes / Q000166:cytology; D003470:Culture Media; D005110:Extracellular Space / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D010477:Perfusion / Q000295:instrumentation*; D012685:Sepharose; D054457:Tissue Scaffolds
chemical_list: D003470:Culture Media; D012685:Sepharose