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.
Cytotechnology. 2005 Jan;47(1-3):3-10. doi: 10.1007/s10616-005-3743-x.
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 mul 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 mum 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.
在这项研究中，我们提出了一种使用涂有 2-（甲基丙烯酰氧）乙基磷酰胆碱（MPC 板）的圆底 96 孔聚苯乙烯板培养小鼠胚胎干细胞（ES 细胞）形成类胚体（EB）的培养方法。MPC 是一种磷脂生物相容性聚合物，可防止细胞黏附在培养表面。将 ES 细胞以每孔 1000 个细胞的密度接种在含有 200ul 培养基的 MPC 板中。经过 5 天的静态孵育，在一个孔中形成了一个称为 EB 的球形细胞聚集体。形成的 EB 的大小（直径）约为 550 微米，包含约 22000 个细胞。似乎非黏附性和孔的圆形是形成良好 EB 的重要因素。将 EB 转移到附着分化培养中，EB 展开并变平，并且在 EB 的生长中有效地产生了搏动细胞（心肌细胞）。涂有 MPC 的圆底 96 孔聚苯乙烯板是形成 EB 的有效工具。
