Dondajewska Ewelina, Juzwa Wojciech, Mackiewicz Andrzej, Dams-Kozlowska Hanna
Chair of Medical Biotechnology, Poznan University of Medical Sciences, Poznan 60-806, Poland.
Department of Biotechnology and Food Microbiology, Poznan University of Life Sciences, 60-627 Poznan, Poland.
Oncotarget. 2017 Dec 22;9(4):4935-4950. doi: 10.18632/oncotarget.23574. eCollection 2018 Jan 12.
An intensive investigation of the development of models to study tumor biology has led to the generation of various three-dimensional (3D) culture methods that better mimic conditions. The tumor microenvironment (TME) is shaped by direct interactions among cancer cells, cancer-associated cells and the extracellular matrix (ECM). Recognizing the need to incorporate both tissue dimensionality and the heterogeneity of cells, we have developed a 3D breast cancer model. NIH3T3 fibroblasts and EMT6 breast cancer cell lines were seeded in various ratios onto a silk fibroin scaffold. The porosity of the silk scaffold was optimized to facilitate the growth of cancer cells. EMT6 and NIH3T3 cells were modified to express GFP and turboFP635, respectively, which enabled the direct analysis of the cell morphology and colonization of the scaffold and for the separation of the cells after their co-culture. Use of 3D mono-culture and 3D co-culture methods resulted in the modification of cell morphology and in a significant increase in ECM production. These culture methods also induced cellular changes related to EMT (epithelial-mesenchymal transition) and CAF (cancer-associated fibroblast) markers. The presented model is an easy to manufacture, well-characterized tool that can be used to study processes of the TME.
对用于研究肿瘤生物学的模型开发进行深入研究，已催生了各种能更好模拟相关条件的三维（3D）培养方法。肿瘤微环境（TME）由癌细胞、癌症相关细胞与细胞外基质（ECM）之间的直接相互作用所塑造。认识到纳入组织维度和细胞异质性的必要性后，我们开发了一种3D乳腺癌模型。将NIH3T3成纤维细胞和EMT6乳腺癌细胞系按不同比例接种到丝素蛋白支架上。对丝支架的孔隙率进行了优化，以促进癌细胞生长。分别对EMT6和NIH3T3细胞进行改造，使其表达绿色荧光蛋白（GFP）和turboFP635，这使得能够直接分析细胞形态以及支架上的细胞定植情况，并在共培养后分离细胞。使用3D单培养和3D共培养方法导致细胞形态发生改变，且细胞外基质产量显著增加。这些培养方法还诱导了与上皮-间质转化（EMT）和癌症相关成纤维细胞（CAF）标志物相关的细胞变化。所展示的模型是一种易于制造、特征明确的工具，可用于研究肿瘤微环境的相关过程。
