人工设计的非接触式多细胞共培养模型可以模仿人体组织的细胞内微环境，但其发展始终面临各种挑战。本研究利用3D打印技术，成功制得了一种含β-磷酸三钙/羟基磷灰石（β-TCP/HA）的细胞容器样支架，该支架设计了四种不同形状的微孔结构：三角形、正方形、平行四边形和长方形微孔结构。这些支架可以在非接触的方式下同时培养四种细胞。本研究构建了一种由人骨髓间质干细胞（HBMSC）、人脐静脉内皮细胞（HUVEC）、人脐静脉平滑肌细胞（HUVSMC）、人真皮成纤维细胞（HDF）组成的3D共培养模型，用以研究这些细胞在促进骨生成和血管生成过程中细胞的个体效应与协同效应。结果表明，相较于在3D细胞容器中仅培养一种细胞，共培养三种或四种细胞展现出了更高的细胞增殖率。HBMSC与HUVEC的细胞间相互作用的研究表明，含有四种独立空间结构的3D细胞容器可以通过放大共培养细胞的旁分泌效应促进细胞的骨生成和血管生成能力。此外，在3D细胞容器中建立多细胞非接触体系，特别是含有三种或四种细胞的共培养体系，相对于单细胞培养模式与两种细胞共培养模式来说，在促进细胞成骨分化和成血管分化方面展现出明显的优势。本研究为基于支架的多细胞非接触式共培养体系的发展提供了新的研究方向。

The development of an engineered non-contact multicellular coculture model that can mimic the in vivo cell microenvironment of human tissues remains challenging. In this study, we successfully fabricated a cell-container-like scaffold composed of β-tricalcium phosphate/hydroxyapatite (β-TCP/HA) bioceramic that contains four different pore structures, including triangles, squares, parallelograms, and rectangles, by means of three-dimensional (3D) printing technology. These scaffolds can be used to simultaneously culture four types of cells in a non-contact way. An engineered 3D coculture model composed of human bone-marrow-derived mesenchymal stem cells (HBMSCs), human umbilical vein endothelial cells (HUVECs), human umbilical vein smooth muscle cells (HUVSMCs), and human dermal fibroblasts (HDFs) with a spatially controlled distribution was constructed to investigate the individual or synergistic effects of these cells in osteogenesis and angiogenesis. The results showed that three or four kinds of cells cocultured in 3D cell containers exhibited a higher cell proliferation rate in comparison with that of a single cell type. Detailed studies into the cell–cell interactions between HBMSCs and HUVECs revealed that the 3D cell containers with four separate spatial structures enhanced the angiogenesis and osteogenesis of cells by amplifying the paracrine effect of the cocultured cells. Furthermore, the establishment of multicellular non-contact systems including three types of cells and four types of cells, respectively, cocultured in 3D cell containers demonstrated obvious advantages in enhancing osteogenic and angiogenic differentiation in comparison with monoculture modes and two-cell coculture modes. This study offers a new direction for developing a scaffold-based multicellular non-contact coculture system for tissue regeneration.