Svanström Andreas, Rosendahl Jennifer, Salerno Simona, Jonasson Emma, Håkansson Joakim, Ståhlberg Anders, Landberg Göran
Department of Laboratory Medicine, Institute of Biomedicine, Sahlgrenska Center for Cancer Research, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
Unit of Biological Function, Division Materials and Production, RISE Research Institutes of Sweden, Borås, Sweden.
Front Bioeng Biotechnol. 2021 Nov 4;9:711977. doi: 10.3389/fbioe.2021.711977. eCollection 2021.
The field of 3D cell cultures is currently emerging, and material development is essential in striving toward mimicking the microenvironment of a native tissue. By using the response of reporter cells to a 3D environment, a comparison between materials can be assessed, allowing optimization of material composition and microenvironment. Of particular interest, the response can be different in a normoxic and hypoxic culturing conditions, which in turn may alter the conclusion regarding a successful recreation of the microenvironment. This study aimed at determining the role of such environments to the conclusion of a better resembling cell culture model to native tissue. Here, the breast cancer cell line MCF7 was cultured in normoxic and hypoxic conditions on patient-derived scaffolds and compared at mRNA and protein levels to cells cultured on 3D printed scaffolds, Matrigel, and conventional 2D plastics. Specifically, a wide range of mRNA targets (40), identified as being regulated upon hypoxia and traditional markers for cell traits (cancer stem cells, epithelial-mesenchymal transition, pluripotency, proliferation, and differentiation), were used together with a selection of corresponding protein targets. 3D cultured cells were vastly different to 2D cultured cells in gene expression and protein levels on the majority of the selected targets in both normoxic and hypoxic culturing conditions. By comparing Matrigel and 3DPS-cultured cells to cells cultured on patient-derived scffolds, differences were also noted along all categories of mRNA targets while specifically for the GLUT3 protein. Overall, cells cultured on patient-derived scaffolds closely resembled cells cultured on 3D printed scaffolds, contrasting 2D and Matrigel-cultured cells, regardless of a normoxic or hypoxic culturing condition. Thus, these data support the use of either a normoxic or hypoxic culturing condition in assays using native tissues as a blueprint to optimize material composition.
3D细胞培养领域目前正在兴起，材料开发对于努力模拟天然组织的微环境至关重要。通过利用报告细胞对3D环境的反应，可以评估材料之间的差异，从而优化材料组成和微环境。特别值得注意的是，在常氧和低氧培养条件下反应可能不同，这反过来可能会改变关于成功重建微环境的结论。本研究旨在确定此类环境在构建更类似于天然组织的细胞培养模型结论中的作用。在这里，乳腺癌细胞系MCF7在常氧和低氧条件下在患者来源的支架上培养，并在mRNA和蛋白质水平上与在3D打印支架、基质胶和传统2D塑料上培养的细胞进行比较。具体而言，一系列被确定在低氧时受到调控的mRNA靶点（40个）以及细胞特征的传统标志物（癌症干细胞、上皮-间质转化、多能性、增殖和分化）与一系列相应的蛋白质靶点一起使用。在常氧和低氧培养条件下，3D培养的细胞在大多数选定靶点的基因表达和蛋白质水平上与2D培养的细胞有很大不同。通过将基质胶和3DPS培养的细胞与在患者来源支架上培养的细胞进行比较，在所有mRNA靶点类别中也发现了差异，特别是对于GLUT3蛋白。总体而言，无论常氧还是低氧培养条件，在患者来源支架上培养的细胞与在3D打印支架上培养的细胞非常相似，这与2D和基质胶培养的细胞形成对比。因此，这些数据支持在以天然组织为蓝图优化材料组成的实验中使用常氧或低氧培养条件。
