Masiello Timothy, Dhall Atul, Hemachandra L P Madhubhani, Tokranova Natalya, Melendez J Andres, Castracane James
Colleges of Nanoscale Science and Engineering, SUNY Polytechnic Institute, Albany, NY 12203, USA.
Cells. 2018 Dec 19;7(12):277. doi: 10.3390/cells7120277.
The transcoelomic metastasis pathway is an alternative to traditional lymphatic/hematogenic metastasis. It is most frequently observed in ovarian cancer, though it has been documented in colon and gastric cancers as well. In transcoelomic metastasis, primary tumor cells are released into the abdominal cavity and form cell aggregates known as spheroids. These spheroids travel through the peritoneal fluid and implant at secondary sites, leading to the formation of new tumor lesions in the peritoneal lining and the organs in the cavity. Models of this process that incorporate the fluid shear stress (FSS) experienced by these spheroids are few, and most have not been fully characterized. Proposed herein is the adaption of a known dynamic cell culture system, the orbital shaker, to create an environment with physiologically-relevant FSS for spheroid formation. Experimental conditions (rotation speed, well size and cell density) were optimized to achieve physiologically-relevant FSS while facilitating the formation of spheroids that are also of a physiologically-relevant size. The FSS improves the roundness and size consistency of spheroids versus equivalent static methods and are even comparable to established high-throughput arrays, while maintaining nearly equivalent viability. This effect was seen in both highly metastatic and modestly metastatic cell lines. The spheroids generated using this technique were fully amenable to functional assays and will allow for better characterization of FSS's effects on metastatic behavior and serve as a drug screening platform. This model can also be built upon in the future by adding more aspects of the peritoneal microenvironment, further enhancing its in vivo relevance.
经体腔转移途径是传统淋巴/血行转移之外的另一种转移方式。它最常见于卵巢癌，不过在结肠癌和胃癌中也有记录。在经体腔转移中，原发性肿瘤细胞释放到腹腔中并形成称为球体的细胞聚集体。这些球体通过腹膜液移动并植入到继发部位，导致在腹膜内衬和腔内器官中形成新的肿瘤病变。纳入这些球体所经历的流体剪切应力（FSS）的该过程模型很少，并且大多数尚未得到充分表征。本文提出的是对一种已知的动态细胞培养系统——轨道振荡器进行改造，以创建一个具有生理相关FSS的球体形成环境。对实验条件（转速、孔大小和细胞密度）进行了优化，以实现生理相关的FSS，同时促进形成生理相关大小的球体。与等效的静态方法相比，FSS提高了球体的圆度和大小一致性，甚至与已建立的高通量阵列相当，同时保持几乎相同的活力。在高转移性和低转移性细胞系中均观察到这种效果。使用该技术产生的球体完全适用于功能测定，将有助于更好地表征FSS对转移行为的影响，并作为药物筛选平台。该模型未来还可以通过增加腹膜微环境的更多方面来进一步构建，进一步增强其体内相关性。
