A 3-D in vitro co-culture model of mammary gland involution
A 3-D in vitro co-culture model of mammary gland involution
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DOI：
10.1039/c3ib40257f
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发表时间：
2014-01-01
影响因子：
2.5
通讯作者：
Watson, Christine J.
中科院分区：
文献类型：
--
作者：
Campbell, Jonathan J.;Botos, Laur-Alexandru;Watson, Christine J.
复归是乳腺在断奶后经历广泛组织重塑的过程，包括细胞死亡、细胞外基质降解和脂肪组织再生。这些过程在一定程度上是通过Jak/Stat信号通路介导的，而Jak/Stat信号通路在乳腺癌中可能被解除调控。体外合成的乳腺类似物可能成为研究肿瘤发生过程的重要工具，或作为个性化药物发现平台和治疗反应的预测因子。理想情况下，这些模型应支持3D新组织形成，以再现生理器官功能，并与高通量筛选方法兼容。我们在工程多孔胶原/透明质酸基质中结合了上皮、基质和免疫来源的细胞系，展示了3d特异性分子特征。此外，种子细胞形成类似乳腺的分支组织，具有小叶肺泡结构，经历诱导内翻表型，使人联想到激素/细胞因子调节下的天然腺体。我们证实，在去除催乳素刺激（H/WD）后的早期时间点，自噬在分化的乳腺上皮细胞内以stat依赖的方式介导。此外，上皮细胞在H/WD下表达M2巨噬细胞谱系的标记物，这一过程随着单核细胞/巨噬细胞系RAW 264.7的引入而减弱。因此，这种3D模型是研究与癌症有关的细胞-细胞相互作用和细胞死亡机制的合适平台。
Involution is a process whereby the mammary gland undergoes extensive tissue remodelling involving exquisitely coordinated cell death, extracellular matrix degradation and adipose tissue regeneration following the weaning of offspring. These processes are mediated in part through Jak/Stat signalling pathways, which can be deregulated in breast cancer. Synthetic in vitro analogues of the breast could become important tools for studying tumorigenic processes, or as personalized drug discovery platforms and predictors of therapeutic response. Ideally, such models should support 3D neo-tissue formation, so as to recapitulate physiological organ function, and be compatible with high-throughput screening methodologies. We have combined cell lines of epithelial, stromal and immunological origin within engineered porous collagen/hyaluronic acid matrices, demonstrating 3D-specific molecular signatures. Furthermore seeded cells form mammary-like branched tissues, with lobuloalveolar structures that undergo inducible involution phenotypes reminiscent of the native gland under hormonal/cytokine regulation. We confirm that autophagy is mediated within differentiated mammary epithelial cells in a Stat-dependent manner at early time points following the removal of a prolactin stimulus (H/WD). In addition, epithelial cells express markers of an M2 macrophage lineage under H/WD, a process that is attenuated with the introduction of the monocyte/macrophage cell line RAW 264.7. Thus, such 3D models are suitable platforms for studying cell-cell interactions and cell death mechanisms in relation to cancer.