A Dynamic Culture Method to Produce Ovarian Cancer Spheroids under Physiologically-Relevant Shear Stress
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 abdomin...
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MDPI AG
2018-12-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/7/12/277 |
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| author | Timothy Masiello Atul Dhall L. P. Madhubhani Hemachandra Natalya Tokranova J. Andres Melendez James Castracane |
| author_facet | Timothy Masiello Atul Dhall L. P. Madhubhani Hemachandra Natalya Tokranova J. Andres Melendez James Castracane |
| author_sort | Timothy Masiello |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-12T09:17:57Z |
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| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-12T09:17:57Z |
| publishDate | 2018-12-01 |
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| series | Cells |
| spelling | doaj.art-60163b9b0e24458083984846cc0bfbe32023-09-02T14:42:35ZengMDPI AGCells2073-44092018-12-0171227710.3390/cells7120277cells7120277A Dynamic Culture Method to Produce Ovarian Cancer Spheroids under Physiologically-Relevant Shear StressTimothy Masiello0Atul Dhall1L. P. Madhubhani Hemachandra2Natalya Tokranova3J. Andres Melendez4James Castracane5Colleges of Nanoscale Science and Engineering, SUNY Polytechnic Institute, Albany, NY 12203, USAColleges of Nanoscale Science and Engineering, SUNY Polytechnic Institute, Albany, NY 12203, USAColleges of Nanoscale Science and Engineering, SUNY Polytechnic Institute, Albany, NY 12203, USAColleges of Nanoscale Science and Engineering, SUNY Polytechnic Institute, Albany, NY 12203, USAColleges of Nanoscale Science and Engineering, SUNY Polytechnic Institute, Albany, NY 12203, USAColleges of Nanoscale Science and Engineering, SUNY Polytechnic Institute, Albany, NY 12203, USAThe 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.https://www.mdpi.com/2073-4409/7/12/277ovariancancerperitonealspheroidshearshakerfluiddynamic |
| spellingShingle | Timothy Masiello Atul Dhall L. P. Madhubhani Hemachandra Natalya Tokranova J. Andres Melendez James Castracane A Dynamic Culture Method to Produce Ovarian Cancer Spheroids under Physiologically-Relevant Shear Stress Cells ovarian cancer peritoneal spheroid shear shaker fluid dynamic |
| title | A Dynamic Culture Method to Produce Ovarian Cancer Spheroids under Physiologically-Relevant Shear Stress |
| title_full | A Dynamic Culture Method to Produce Ovarian Cancer Spheroids under Physiologically-Relevant Shear Stress |
| title_fullStr | A Dynamic Culture Method to Produce Ovarian Cancer Spheroids under Physiologically-Relevant Shear Stress |
| title_full_unstemmed | A Dynamic Culture Method to Produce Ovarian Cancer Spheroids under Physiologically-Relevant Shear Stress |
| title_short | A Dynamic Culture Method to Produce Ovarian Cancer Spheroids under Physiologically-Relevant Shear Stress |
| title_sort | dynamic culture method to produce ovarian cancer spheroids under physiologically relevant shear stress |
| topic | ovarian cancer peritoneal spheroid shear shaker fluid dynamic |
| url | https://www.mdpi.com/2073-4409/7/12/277 |
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