A Round-bottom 96-well Polystyrene Plate Coated with 2-methacryloyloxyethyl Phosphorylcholine as an Effective Tool for Embryoid Body Formation |
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Authors: | Mikiko Koike Hiroshi Kurosawa Yoshifumi Amano |
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Institution: | (1) Life Environment Medical Engineering, Division of Medicine and Engineering Science, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, 4-3-11 Takeda, Kofu Yamanashi 400-8511, Japan |
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Abstract: | In this study, we proposed a culture method for forming embryoid bodies (EBs) from mouse embryonic stem (ES) cells using a
round-bottom 96-well polystyrene plate coated with 2-methacryloyloxyethyl phosphorylcholine (MPC plate). MPC is a phospholipid
biocompatible polymer and prevents cells from adhering to the culture surface. The ES cells were seeded at 1000 cells per
well in the MPC plate with 200 μl of medium. After 5 days of static incubation, a spherical cell aggregate termed EB was formed
in a well. The size (diameter) of resulting EB was approximately 550 μm and it contained approx. 22,000 cells. It seems that
the non-adhesiveness and the roundness of the well are important factors to form a good EB. Transferring the EBs to the attached
differentiation culture, the EBs spread out and flattened, and the beating cells (cardiomyocytes) were effectively generated
in the outgrowth of EBs. The round-bottom 96-well polystyrene plate coated with MPC is an effective tool for EB formation. |
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Keywords: | Differentiation Embryoid body (EB) ES cell Methacryloyloxyethyl phosphorylcholine (MPC) Non-adhesiveness |
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