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Microfluidic perfusion culture of human induced pluripotent stem cells under fully defined culture conditions
Author(s) -
Yoshimitsu Ryosuke,
Hattori Koji,
Sugiura Shinji,
Kondo Yuki,
Yamada Rotaro,
Tachikawa Saoko,
Satoh Taku,
Kurisaki Akira,
Ohnuma Kiyoshi,
Asashima Makoto,
Kanamori Toshiyuki
Publication year - 2014
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.25150
Subject(s) - induced pluripotent stem cell , human induced pluripotent stem cells , cell culture , microfluidics , stem cell , microbiology and biotechnology , biology , chemistry , nanotechnology , materials science , embryonic stem cell , biochemistry , genetics , gene
Human induced pluripotent stem cells (hiPSCs) are a promising cell source for drug screening. For this application, self‐renewal or differentiation of the cells is required, and undefined factors in the culture conditions are not desirable. Microfluidic perfusion culture allows the production of small volume cultures with precisely controlled microenvironments, and is applicable to high‐throughput cellular environment screening. Here, we developed a microfluidic perfusion culture system for hiPSCs that uses a microchamber array chip under defined extracellular matrix (ECM) and culture medium conditions. By screening various ECMs we determined that fibronectin and laminin are appropriate for microfluidic devices made out of the most popular material, polydimethylsiloxane (PDMS). We found that the growth rate of hiPSCs under pressure‐driven perfusion culture conditions was higher than under static culture conditions in the microchamber array. We applied our new system to self‐renewal and differentiation cultures of hiPSCs, and immunocytochemical analysis showed that the state of the hiPSCs was successfully controlled. The effects of three antitumor drugs on hiPSCs were comparable between microchamber array and 96‐well plates. We believe that our system will be a platform technology for future large‐scale screening of fully defined conditions for differentiation cultures on integrated microfluidic devices. Biotechnol. Biotechnol. Bioeng. 2014;111: 937–947. © 2013 Wiley Periodicals, Inc.