Koch Maria K, Jaeschke Anna, Murekatete Berline, Ravichandran Akhilandeshwari, Tsurkan Mikhail, Werner Carsten, Soon Patsy, Hutmacher Dietmar W, Haupt Larisa M, Bray Laura J
Institute of Health and Biomedical Innovation, Queensland University of Technology, Australia; School of Mechanical, Medical and Process Engineering, Science and Engineering Faculty, Queensland University of Technology, 60 Musk Avenue, Kelvin Grove, QLD 4059, Australia.
Institute of Health and Biomedical Innovation, Queensland University of Technology, Australia.
Acta Biomater. 2020 Sep 15;114:256-269. doi: 10.1016/j.actbio.2020.07.036. Epub 2020 Jul 21.
The plasticity of the tumour microenvironment is a key contributor to cancer development and progression. Here, we present a bioengineered breast tumour angiogenesis model comprised of mammary derived epithelial, endothelial and fibroblast cells, to dissect the mechanisms of cancer-associated fibroblasts (CAFs) on microvascular-like network formation and epithelial spheroid morphology. Primary patient-derived mammary endothelial cells, normal breast fibroblasts (NBF, patient matched) and CAFs were cultured within three-dimensional (3D) semi-synthetic hydrogels where CAFs promoted an increase in the density and morphology of the microvascular-like network. The mammary microenvironment also increased the number of MCF-10a epithelial spheroids when compared with a non-mammary microenvironment, and a malignant mammary microenvironment resulted in further morphological differences in the epithelial spheroids. The morphological changes observed following interactions between breast CAFs and endothelial cells, highlight the plasticity of the malignant stroma in tumour vascularisation. Our in vitro bioengineered breast cancer microenvironment provides a robust model to study cell-cell and cell-matrix interactions. Statement of Significance In recent years there has been an increase in the sophistication of 3D culture models, however less attention has been paid to the cell source utilised. In this study, we describe the influence of a normal and malignant stromal microenvironment on vessel-like behaviour in a 3D model. Using a semi-synthetic hydrogel, we studied the effects of mammary-derived cancer-associated fibroblasts and normal fibroblasts on human umbilical vein endothelial cells or human mammary microvascular endothelial cells. An increase in vessel-like network and epithelial cell density was seen in a mammary versus non-mammary microenvironment. This study highlights the importance of using tissue-specific endothelial cells in cancer research and demonstrates the microenvironmental impact of fibroblasts on endothelial and epithelial growth and morphology.
肿瘤微环境的可塑性是癌症发生和发展的关键因素。在此，我们展示了一种生物工程化的乳腺肿瘤血管生成模型，该模型由乳腺来源的上皮细胞、内皮细胞和成纤维细胞组成，用于剖析癌症相关成纤维细胞（CAFs）对微血管样网络形成和上皮球状体形态的作用机制。将原发性患者来源的乳腺内皮细胞、正常乳腺成纤维细胞（NBF，与患者匹配）和CAFs培养在三维（3D）半合成水凝胶中，其中CAFs促进了微血管样网络密度和形态的增加。与非乳腺微环境相比，乳腺微环境也增加了MCF - 10a上皮球状体的数量，而恶性乳腺微环境导致上皮球状体出现进一步的形态差异。乳腺CAFs与内皮细胞相互作用后观察到的形态变化，突出了恶性基质在肿瘤血管生成中的可塑性。我们的体外生物工程化乳腺癌微环境为研究细胞 - 细胞和细胞 - 基质相互作用提供了一个强大的模型。意义声明近年来，3D培养模型越来越复杂，但对所使用的细胞来源关注较少。在本研究中，我们描述了正常和恶性基质微环境对3D模型中血管样行为的影响。使用半合成水凝胶，我们研究了乳腺来源的癌症相关成纤维细胞和正常成纤维细胞对人脐静脉内皮细胞或人乳腺微血管内皮细胞的影响。与非乳腺微环境相比，乳腺微环境中血管样网络和上皮细胞密度增加。本研究强调了在癌症研究中使用组织特异性内皮细胞的重要性，并证明了成纤维细胞对内皮细胞和上皮细胞生长及形态的微环境影响。
