Microfluidic perfusion culture of human induced pluripotent stem cells under fully defined culture conditions.
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.
Department of Bioengineering, Nagaoka University of Technology, 1603-1 Kamitomioka, Nagaoka, Niigata, 940-2188, Japan.
Ryosuke
R
Yoshimitsu
Koji
K
Hattori
Shinji
S
Sugiura
Yuki
Y
Kondo
Rotaro
R
Yamada
Saoko
S
Tachikawa
Taku
T
Satoh
Akira
A
Kurisaki
Kiyoshi
K
Ohnuma
Makoto
M
Asashima
Toshiyuki
T
Kanamori
D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002454:Cell Differentiation; D004867:Equipment Design; D016326:Extracellular Matrix Proteins; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D010477:Perfusion / Q000295:instrumentation* / Q000379:methods; D039904:Pluripotent Stem Cells / Q000166:cytology*
D016326:Extracellular Matrix Proteins
cell-based assay; defined culture conditions; human induced pluripotent stem cells; microchamber array; monolayer culture; perfusion culture