Embryoid body culture of mouse embryonic stem cells using microwell and micropatterned chips(CELL AND TISSUE ENGINEERING)
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概要
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The proliferation and differentiation properties of embryoid bodies (EB) from mouse embryonic stem (ES) cells were compared under two microchip conditions: microwell chip and micropatterned chip. The microwell chip contained 270 microwells (diameter, 600μm; depth, 600μm) on a polymethylmethacrylate plate and was surface-modified with polyethylene glycol (PEG) to render it non-adhesive. The micropatterned chip contained 270 gelatin spots (diameter, 200μm) as the cell adhesion area on a glass plate; the region lacking these spots was PEG-modified to render it non-adhesive. The ES cells spontaneously formed the EBs from cell aggregates in each microwell in the chip. In contrast, cells inoculated onto the patterned chip formed a monolayer on the gelatin spots and gradually proliferated to form EBs. The EBs in the patterned chip maintained the high cell growth rate and the expression of endoderm (TTR and AFP) and mesoderm (Nkx2.5, αMHC, Flk1, and PDGFRβ) markers was increased, and these cell properties were similar to the previous methods (hanging drop and round-bottomed 96-well plate cultures). In contrast, the proliferation of ES cells in the microwell chip was lower than in the patterned chip and previous methods, and the EB differentiation proceeded slowly and only formed a small amount of endoderm. These results indicate that the difference of EB generating process in the microchip cultures may affect to the proliferation and differentiation of ES cells, and the existence of microwell structure in the microchip downregulates the cell proliferation and the differentiated progress of ES cells.
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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Yoshiura Yukiko
Department of Life and Environment Engineering, The University of Kitakyushu
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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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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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Sakai Yusuke
Department Of Energy Sciences Tokyo Institute Of Technology
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Yoshiura Yukiko
Department Of Life And Environment Engineering The University Of Kitakyushu
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