The Biophysical Interdisciplinary Jerome Schottenstein Center for the Research and Technology of the Cellome, Physics Department, Bar-Ilan University, Ramat-Gan 52900, Israel.
The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan 52900, Israel.
Toxicol In Vitro. 2021 Mar;71:105067. doi: 10.1016/j.tiv.2020.105067. Epub 2020 Dec 8.
In response to the need for reliable cellular models that reflect complex tumor microenvironmental properties, and enable more precise testing of anti-cancer therapeutics effects on humans, a co-culture platform for in-vitro model that enhances the physiology of breast cancer (BC) microenvironment is presented. A six well imaging plate wherein each macro-well contains several separate compartments was designed. Three-dimensional (3D) cancer spheroids are generated and cultured in the inner compartment which is embossed with an array of nano-liter micro-chambers made of hydrogel. Stromal cells are cultured in the outer chambers. The two cell types are cultured side-by-side, sharing a common space, thus enabling extra-cellular communication via secreted molecules. As proof of concept, a model of BC tumor microenvironment was recapitulated by co-cultivating 3D MCF7 spheroids in the presence of tumor-associated macrophages (TAMs). The presence of TAMs induced an aggressive phenotype by promoting spheroid growth, enhancing survivin expression levels and enabling invasive behavior. Moreover, TAMs influenced the response of BC spheroids to cytotoxic treatment as well as hormonal drug therapy, and enhanced the effects of nitric oxide donor. The platform enables time-lapse imaging and treatment without losing spatial location of the measured spheroids, thereby allowing measurements and analysis at individual-object resolution in an easy and efficient manner.
为了满足对能够反映复杂肿瘤微环境特性并能够更精确地测试抗癌疗法对人类影响的可靠细胞模型的需求，本文提出了一种体外模型共培养平台，以增强乳腺癌（BC）微环境的生理学特性。设计了一个六孔成像板，其中每个大孔包含几个单独的隔室。三维（3D）癌症球体在内部隔室中生成和培养，该隔室压印有一系列由水凝胶制成的纳升微室阵列。基质细胞在外部隔室中培养。两种细胞类型并排培养，共享一个共同的空间，从而通过分泌的分子实现细胞外通讯。作为概念验证，通过共培养存在肿瘤相关巨噬细胞（TAMs）的 3D MCF7 球体，再现了 BC 肿瘤微环境模型。TAMs 的存在通过促进球体生长、增强生存素表达水平和促进侵袭行为，诱导侵袭表型。此外，TAMs 还影响 BC 球体对细胞毒性治疗和激素药物治疗的反应，并增强了一氧化氮供体的作用。该平台允许在不丢失测量球体空间位置的情况下进行延时成像和治疗，从而能够以简单有效的方式在单个对象分辨率下进行测量和分析。
