A round-bottom 96-well polystyrene plate coated with 2-methacryloyloxyethyl phosphorylcholine as an effective tool for embryoid body formation
A round-bottom 96-well polystyrene plate coated with 2-methacryloyloxyethyl phosphorylcholine as an effective tool for embryoid body formation
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DOI：
10.1007/s10616-005-3743-x
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发表时间：
2005-01-01
期刊：
影响因子：
2.2
通讯作者：
Amano, Y
中科院分区：
文献类型：
--
作者：
Koike, M;Kurosawa, H;Amano, Y
本研究提出了一种用96孔聚苯乙烯圆底2-甲基丙烯酰氧乙基磷酸胆碱（MPC）板培养小鼠胚胎干细胞形成胚状体（EBs）的方法。MPC是一种磷脂生物相容性聚合物，可防止细胞粘附在培养表面。将胚胎干细胞以每孔1000个细胞的速度接种在MPC板上，培养基为200 μ l。静孵育5天后，在井中形成称为EB的球形细胞聚集体。所产生的EB的大小（直径）约为550 μ m，其中包含约。22000个细胞。不粘接性和井的圆度是形成良好EB的重要因素。将EBs转移到附着的分化培养物中，EBs展开并变平，并且在EBs的生长中有效地产生了跳动细胞（心肌细胞）。涂覆MPC的96孔聚苯乙烯圆底板是形成EB的有效工具。
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 mu l 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 mu m 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.