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Plasma Medicine
SJR: 0.278 SNIP: 0.183 CiteScore™: 0.57

ISSN Print: 1947-5764
ISSN Online: 1947-5772

Plasma Medicine

DOI: 10.1615/PlasmaMed.2014012077
pages 165-176

Plasma-Patterned Polydimethylsiloxane Surface With Single-Step Coating with a Mixture of Vitronectin and Albumin Enables the Formation of Small Discs and Spheroids of Human Induced Pluripotent Stem Cells

Ryotaro Yamada
Department of Bioengineering, Nagaoka University of Technology, Nagaoka, Niigata, Japan
Koji Hattori
Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan
Motohiro Tagaya
Department of Materials Science and Technology, Nagaoka University of Technology, Nagaoka, Niigata, Japan
Toru Sasaki
Department of Electrical Engineering, Nagaoka University of Technology, Nagaoka, Niigata, Japan
Daisuke Miyamoto
Faculty of Environmental Engineering, The University of Kitakyushu, Kitakyushu, Fukuoka, Japan
Kohji Nakazawa
Faculty of Environmental Engineering, The University of Kitakyushu, Kitakyushu, Fukuoka, Japan
Shinji Sugiura
Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan
Toshiyuki Kanamori
Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan
Kiyoshi Ohnuma
Department of Bioengineering, Nagaoka University of Technology, Nagaoka, Niigata, Japan

ABSTRACT

Plasma treatment is an easy method of cleaning and hydrophilizing glass and polymer surfaces to facilitate adsorption of cell-adhesive proteins. Thus a patterned plasma treatment is useful for fabricating cell adhesion patterns. We previously succeeded in producing human induced pluripotent stem cell (hiPSC) discs (2 mm in diameter) on plasma-patterned polydimethylsiloxane (PDMS) by single-step coating of a mixture of 2 ubiquitous proteins, vitronectin and γ-globulin. Under serum-free and feeder-free conditions, without any undefined cell adhesion molecules, γ-globulin blocked hiPSC adhesion on surfaces not treated with plasma. However, γ-globulin has low cost-effectiveness and availability, and the resulting disc diameter was too large for cell-based assays. We demonstrate that bovine serum albumin (BSA) can also be used to block hiPSC adhesion on plasma-untreated PDMS surfaces coated with vitronectin. We succeeded in creating small hiPSC discs (200 µm in diameter) using single-step coating of a mixture of vitronectin and BSA. The hiPSCs proliferated without escaping from the patterned area and finally spontaneously detached from the discs to form spheroids. We believe that our method for generating hiPSC discs and spheroids will be useful for developing new bioengineering devices to enhance cell differentiation and to test drug safety for human embryonic development, contributing to future medical applications.


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