Development of temperature-responsive culture dishes which allow viable cell sheet recovery

多种细胞类型的共培养是体外组织构建的关键前提，通常会导致一种细胞类型主导共培养细胞群体。自然组织经常被观察到由多种细胞类型组成，在层状结构中形成不同的层。在不同的器官和组织中，几种细胞类型被共同组织成确定的细胞层，其中细胞层的分层对于细胞与细胞的相互作用以及在更大的水平上对器官的功能至关重要。我们挑战了一种从细胞片重建组织结构的新策略。独特的温度响应型培养表面允许将培养的各种类型的细胞收获为融合的、完整的细胞单层片，而无需典型的破坏性酶处理。覆盖两层单层肝细胞和内皮细胞为体外构建肝小叶样组织提供了一种可行的结构。培养的板层细胞片保留了每个细胞的表型和基本细胞功能。这一通用策略在细胞-细胞相互作用、组织新生和许多类型的组织工程生物材料的设计中具有实用价值。

Co-culture of multiple cell types, a critical pre-requisite for in vitro tissue construction, often results in domination of co-culture cell populations by one cell type. Natural tissues are often observed to comprise multiple cell types forming distinct layers in stratified architectures. Several cell types are co-organized into defined cell layers in various organs and tissues where stratification of cell layers is critical to cell-cell interaction and, at a larger level, to organ function. We have challenged a novel strategy to reconstruct tissue architectures from cell sheets. A unique temperature-responsive culture surface permits harvest of cultured various cell types as confluent, intact cell monolayer sheets without typical destructive enzymatic treatment. Overlaying two monolayer sheets of hepatocytes and endothelial cells provides a viable construct for in vitro fabricated liver lobule-like tissue. Cultured lamellar cell sheets preserve each cell phenotype and basic cell functions. This general strategy has utility in studies of cell-cell interactions, tissue neogenesis and the design of many types of tissue-engineered biomaterials.