Multicellular spheroids represent a well-established 3D model to study healthy and diseased cells in vitro. The use of conventional 3D cell culture platforms for the generation of multicellular spheroids is limited to cell types that easily self-assemble into spheroids because less adhesive cells fail to form stable aggregates. A high-precision micromoulding technique developed in our laboratory produces deep conical agarose microwell arrays that allow the cultivation of uniform multicellular aggregates, irrespective of the spheroid formation capacity of the cells. Such hydrogel arrays warrant a steady nutrient supply for several weeks, permit live volumetric measurements to monitor cell growth, enable immunohistochemical staining, fluorescence-based microscopy, and facilitate immediate harvesting of cell aggregates. This system also allows co-cultures of two distinct cell types either in direct cell–cell contact or at a distance as the hydrogel permits diffusion of soluble compounds. Notably, we show that co-culture of a breast cancer cell line with bone marrow stromal cells enhances 3D growth of the cancer cells in this system.

中文翻译：

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深锥形琼脂糖微孔阵列，用于独立于粘附的三维细胞培养和动态体积测量†

多细胞球体代表了一种完善的3D模型，可以在体外研究健康和患病的细胞。使用常规的3D细胞培养平台生成多细胞球体仅限于容易自组装成球体的细胞类型，因为较少的粘附细胞无法形成稳定的聚集体。在我们实验室中开发的高精度微成型技术可产生深锥形琼脂糖微孔阵列，无论细胞的球状形成能力如何，均可培养均匀的多细胞聚集体。这种水凝胶阵列可保证数周稳定的营养供应，可进行实时体积测量以监测细胞生长，启用免疫组织化学染色，基于荧光的显微镜检查，并有助于立即收集细胞聚集体。由于水凝胶允许可溶性化合物扩散，该系统还允许两种不同细胞类型的共培养进行直接细胞间接触或相隔一定距离。值得注意的是，我们显示乳腺癌细胞系与骨髓基质细胞的共培养可增强该系统中癌细胞的3D生长。