Abstract
Abstract Three-dimensional (3D) cancer cell cultures are being utilized for drug screening with increasing frequency since they provide a more physiologically relevant solid tumor model than two-dimensional cell cultures. Spheroids are a well characterized, in vitro tumor model system that has been utilized with multiple cancer cell lines. Growth and manipulation of these cultures in hanging drop plates (HDPs) can be challenging to manipulate and these challenges are amplified as sample throughput increases. We automated the culture and compound treatment of cancer spheroids in HDPs, as well as the preparation of these spheroids for analysis by high content imaging and flow cytometry. Automated cancer spheroid cultures showed excellent consistency in size and shape, providing higher confidence in the screening results. Creating single cell suspensions from the cancer spheroids for flow cytometry analysis was another significant challenge overcome through automation. Imaging provided information such as the location of drug-responding cells within the cancer spheroid while flow cytometry removed the potential for spheroid exclusion of staining reagents leading to false negatives. Automation enabled the use of these complementary analyses, thereby achieving a more complete understanding of compound sensitivity in solid tumors and illustrating how spheroids can be utilized to screen for novel cancer therapeutics. Citation Format: Michael Kowalski, Vipat Raksakulthai, Kristin Prasauckas, Tara Jones-Roe. Automation-enabled 3D tumor chemosensitivity screening by imaging and flow cytometry analysis. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4265.
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