Effect of microwell chip structure on cell microsphere production of various animal cells(CELL AND TISSUE ENGINEERING)
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概要
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The formation of three-dimensional cell microspheres such as spheroids, embryoid bodies, and neurospheres has attracted attention as a useful culture technique. In this study, we investigated a technique for effective cell microsphere production by using specially prepared microchip. The basic chip design was a multimicrowell structure in triangular arrangement within a 100-mm^2 region in the center of a polymethylmethacrylate (PMMA) plate (24×24mm^2), the surface of which was modified with polyethylene glycol (PEG) to render it nonadhesive to cells. We also designed six similar chips with microwell diameters of 200, 300, 400, 600, 800, and 1000μm to investigate the effect of the microwell diameter on the cell microsphere diameter. Rat hepatocytes, HepG2 cells, mouse embryonic stem (ES) cells, and mouse neural progenitor/stem (NPS) cells formed hepatocyte spheroids, HepG2 spheroids, embryoid bodies, and neurospheres, respectively, in the microwells within 5 days of culture. For all the cells, a single microsphere was formed in each microwell under all the chip conditions, and such microsphere configurations remained throughout the culture period. Furthermore, the microsphere diameters of each type of cell were strongly positively correlated with the microwell diameters of the chips, suggesting that microsphere diameter can be factitiously controlled by using different chip conditions. Thus, this chip technique is a promising cellular platform for tissue engineering or regenerative medicine research, pharmacological and toxicological studies, and fundamental studies in cell biology.
著者
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Sakai Yusuke
Department of Life and Environment Engineering, The University of Kitakyushu
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Nakazawa Kohji
Department of Life and Environment Engineering, The University of Kitakyushu
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Kanemura Yonehiro
Department of Regenerative Medicine, Institute for Clinical Research, Osaka National Hospital, Natio
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Mori Hideki
Department of Biological Science, Graduate School of Science, Osaka Prefecture University
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Yoshida Shirou
Department of Life and Environment Engineering, The University of Kitakyushu
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Yoshiura Yukiko
Department of Life and Environment Engineering, The University of Kitakyushu
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Mori Rhuhei
Department of Life and Environment Engineering, The University of Kitakyushu
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Tamura Tomoko
STEM Biomethod Corporation
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Yahiro Kanji
STEM Biomethod Corporation
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Yamasaki Mami
Department of Neurosurgery, Osaka National Hospital, National Hospital Organization
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Nakazawa Kohji
Department Of Chemical Processes And Environments The University Of Kitakyushu
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Nakazawa Kohji
Department Of Life And Environment Engineering The University Of Kitakyushu
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Nakazawa Kohji
Dep. Of Life And Environment Engineering The Univ. Of Kitakyushu 1-1 Hibikino Wakamatsu-ku Kitakyush
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Mori Hideki
Dep. Of Biological Sci. Graduate School Of Sci. Osaka Prefecture Univ. 1-2 Gakuen-cho Naka-ku Sakai
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Yamasaki Mami
Department Of Neurosurgery Osaka National Hospital National Hospital Organization:department Of Mole
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Yamasaki Mami
Department Of Neurosurgery And Clinical Institute Osaka National Hospital National Hospital Organiza
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Kanemura Yonehiro
Stem Biomethod Corporation
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Sakai Yusuke
Dep. Of Life And Environment Engineering The Univ. Of Kitakyushu 1-1 Hibikino Wakamatsu-ku Kitakyush
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Ninomiya Kazuaki
Department Of Biotechnology Graduate School Of Engineering Osaka University:(present Office)institut
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Ninomiya K
Research Institute For Cell Engineering National Institute Of Advanced Industrial Science And Techno
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Kanemura Yonehiro
Institute For Clinical Research Osaka National Hospital National Hospital Organization:research Inst
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Mori Rhuhei
Department Of Life And Environment Engineering The University Of Kitakyushu
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Mori Hideki
Department Of Anatomy Okayama University Medical School
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Sakai Yusuke
Department Of Energy Sciences Tokyo Institute Of Technology
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Yoshida Shirou
Department Of Life And Environment Engineering The University Of Kitakyushu
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Kanemura Yonehiro
Department Of Regenerative Medicine Institute For Clinical Research Osaka National Hospital National
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Yoshiura Yukiko
Department Of Life And Environment Engineering The University Of Kitakyushu
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Mori Hideki
Division Of Chemical Engineering Graduate School Of Engineering Science Osaka University:institute F
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Mori Hideki
Inst. For Clinical Res. Osaka National Hospital National Hospital Organization
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