Detachably assembled microfluidic device for perfusion culture and post-culture analysis of a spheroid array

Yusuke Sakai, Koji Hattori, Fumiki Yanagawa, Shinji Sugiura, Toshiyuki Kanamori, Kohji Nakazawa

Research output: Contribution to journalArticlepeer-review

25 Citations (Scopus)


Microfluidic devices permit perfusion culture of three-dimensional (3D) tissue, mimicking the flow of blood in vascularized 3D tissue in our body. Here, we report a microfluidic device composed of a two-part microfluidic chamber chip and multi-microwell array chip able to be disassembled at the culture endpoint. Within the microfluidic chamber, an array of 3D tissue aggregates (spheroids) can be formed and cultured under perfusion. Subsequently, detailed post-culture analysis of the spheroids collected from the disassembled device can be performed. This device facilitates uniform spheroid formation, growth analysis in a high-throughput format, controlled proliferation via perfusion flow rate, and post-culture analysis of spheroids. We used the device to culture spheroids of human hepatocellular carcinoma (HepG2) cells under two controlled perfusion flow rates. HepG2 spheroids exhibited greater cell growth at higher perfusion flow rates than at lower perfusion flow rates, and exhibited different metabolic activity and mRNA and protein expression under the different flow rate conditions. These results show the potential of perfusion culture to precisely control the culture environment in microfluidic devices. The construction of spheroid array chambers allows multiple culture conditions to be tested simultaneously, with potential applications in toxicity and drug screening.
Original languageEnglish
Pages (from-to)971-979
Number of pages9
JournalBiotechnology Journal
Issue number7
Publication statusPublished - Jul 2014


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