RRC ID 44735
Author Yoshimitsu R, Hattori K, Sugiura S, Kondo Y, Yamada R, Tachikawa S, Satoh T, Kurisaki A, Ohnuma K, Asashima M, Kanamori T.
Title Microfluidic perfusion culture of human induced pluripotent stem cells under fully defined culture conditions.
Journal Biotechnol Bioeng
Abstract 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.
Volume 111(5)
Pages 937-47
Published 2014-5-1
DOI 10.1002/bit.25150
PMID 24222619
MeSH Cell Culture Techniques / instrumentation* Cell Culture Techniques / methods Cell Differentiation Equipment Design Extracellular Matrix Proteins Humans Microfluidic Analytical Techniques / instrumentation* Microfluidic Analytical Techniques / methods Perfusion / instrumentation* Perfusion / methods Pluripotent Stem Cells / cytology*
IF 4.002
Times Cited 25
WOS Category BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Resource
Human and Animal Cells 201B7(HPS0063)