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先决条件，通常会导致共培养细胞群被一种细胞类型所统治。通常观察到，自然组织包括多种细胞类型，在分层结构中形成不同的层。在各种器官和组织中，几种细胞类型共同组织成确定的细胞层，其中细胞层的分层对细胞间相互作用至关重要，在更大的水平上，对器官功能至关重要。我们挑战了一种从细胞薄片重建组织结构的新策略。独特的温度响应培养表面允许收获培养的各种细胞类型作为融合，完整的细胞单层片，而无需典型的破坏性酶处理。肝细胞和内皮细胞覆盖两层单层片，为体外构建肝小叶样组织提供了可行的结构。培养的层状细胞保留了每个细胞的表型和基本细胞功能。这一总体策略在细胞-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研究中具有实用价值。

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.