Department of Applied Chemistry, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo 113-8656, Japan.
Biomaterials. 2010 Dec;31(34):8839-46. doi: 10.1016/j.biomaterials.2010.07.106. Epub 2010 Aug 21.
The biological performances of a cell-containing phospholipid polymer hydrogel in bulk and miniaturized formats without an additional culture medium support were investigated and compared. The cell-containing hydrogel was formed spontaneously when solutions of commercial polyvinyl alcohol (PVA) and the phospholipid polymer poly[2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate (BMA)-co-p-vinylphenylboronic acid (VPBA)] (PMBV) suspended with cells in a cell culture medium are mixed together. Bulk and miniaturized hydrogels, with approximate thicknesses of 3.1 mm and 400 μm, respectively, were prepared in a 96-well microplate and a glass microchip, respectively. In both cases, the hydrogels were homogeneous, and cells were spatially encapsulated. The long-term observation (4 and 8 days) of cell morphology suggested that cells were passively attached to the interface of the hydrogel but were unable to spread and flatten, which inhibited cell growth in both hydrogels. Viability evaluations revealed that cells in both hydrogel formats maintained the same high viability levels after long-term encapsulation. Cytotoxicity assays indicated that the cells in the miniaturized hydrogel maintained a high degree of correlation in cytotoxic sensitivity with the cells in the bulk hydrogel and a routine medium culture. The PMBV/PVA hydrogel not only provides a beneficial cytocompatible microenvironment for long-term cell survival without an additional culture medium support but also creates a static condition for cell sustainment in a microchip similar to that in bulk. The uniform long-term performances of PMBV/PVA hydrogels in bulk and miniaturized formats make them ideal for the development of long-term, flexible, three-dimensional, living cell-based tools for routine cell-based assays and applications on bulk to microscale levels.
研究并比较了在没有额外培养基支持的情况下,含有细胞的磷脂聚合物水凝胶在块状和微型化形式中的生物学性能。当含有细胞的商业聚乙烯醇(PVA)和磷脂聚合物聚[2-甲基丙烯酰氧基乙基磷酸胆碱(MPC)-共-正丁基甲基丙烯酸酯(BMA)-共-p-乙烯基苯硼酸(VPBA)](PMBV)溶液与悬浮在细胞培养基中的细胞混合时,会自发形成含细胞的水凝胶。分别在 96 孔微孔板和玻璃微芯片中制备了厚度约为 3.1mm 和 400μm 的块状和微型水凝胶。在这两种情况下,水凝胶都是均匀的,并且细胞被空间包封。细胞形态的长期观察(4 和 8 天)表明,细胞被动地附着在水凝胶的界面上,但无法扩散和平整,这抑制了两种水凝胶中的细胞生长。活力评估表明,长期包封后,两种水凝胶格式中的细胞均保持相同的高活力水平。细胞毒性测定表明,微型水凝胶中的细胞在细胞毒性敏感性方面与大块水凝胶和常规培养基培养中的细胞高度相关。PMBV/PVA 水凝胶不仅在没有额外培养基支持的情况下为细胞的长期生存提供了有益的细胞相容性微环境,而且在类似于大块的微芯片中为细胞的维持创造了静态条件。PMBV/PVA 水凝胶在块状和微型化形式中的均匀长期性能使其成为常规基于细胞的测定和从大块到微尺度水平的基于细胞的应用的长期、灵活、三维、活细胞工具的理想选择。