Current materials used for in vitro 3D cell culture are often limited by their poor similarity to human tissue, batch-to-batch variability and complexity of composition and manufacture. Here, we present a "blank slate" culture environment based on a self-assembling peptide gel free from matrix motifs. The gel can be customised by incorporating matrix components selected to match the target tissue, with independent control of mechanical properties. Therefore the matrix components are restricted to those specifically added, or those synthesised by encapsulated cells. The flexible 3D culture platform provides full control over biochemical and physical properties, allowing the impact of biochemical composition and tissue mechanics to be separately evaluated in vitro. Here, we demonstrate that the peptide gels support the growth of a range of cells including human induced pluripotent stem cells and human cancer cell lines. Furthermore, we present proof-of-concept that the peptide gels can be used to build disease-relevant models. Controlling the peptide gelator concentration allows peptide gel stiffness to be matched to normal breast (<1 kPa) or breast tumour tissue (>1 kPa), with higher stiffness favouring the viability of breast cancer cells over normal breast cells. In parallel, the peptide gels may be modified with matrix components relevant to human breast, such as collagen I and hyaluronan. The choice and concentration of these additions affect the size, shape and organisation of breast epithelial cell structures formed in co-culture with fibroblasts. This system therefore provides a means of unravelling the individual influences of matrix, mechanical properties and cell-cell interactions in cancer and other diseases.

中文翻译：

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具有完全确定的成分和力学的肽凝胶，用于体外探测细胞-细胞和细胞-基质相互作用。


目前用于体外 3D 细胞培养的材料通常受到与人体组织相似性差、批次间差异以及成分和制造复杂性的限制。在这里，我们提出了一种基于不含基质基序的自组装肽凝胶的“白板”培养环境。凝胶可以通过加入与目标组织相匹配的基质成分来定制，并独立控制机械性能。因此，基质成分仅限于那些专门添加的成分，或者那些由封装细胞合成的成分。灵活的 3D 培养平台可完全控制生化和物理特性，从而可以在体外单独评估生化成分和组织力学的影响。在这里，我们证明肽凝胶支持一系列细胞的生长，包括人类诱导多能干细胞和人类癌细胞系。此外，我们提出了肽凝胶可用于构建疾病相关模型的概念验证。控制肽凝胶剂浓度可以使肽凝胶硬度与正常乳腺相匹配(<1 id=30>1 kPa)，较高的硬度有利于乳腺癌细胞比正常乳腺细胞的活力。同时，肽凝胶可以用与人类乳房相关的基质成分（例如胶原蛋白 I 和乙酰透明质酸）进行修饰。这些添加物的选择和浓度会影响与成纤维细胞共培养时形成的乳腺上皮细胞结构的大小、形状和组织。因此，该系统提供了一种揭示癌症和其他疾病中基质、机械性能和细胞间相互作用的个体影响的方法。