Abstract 208: A novel cell culture surface supports effective formation of three dimensional cancer spheroids in suspension.
Authors: Neeley Cindy , Gaarn Louise , Marwood Tina , Scott Robert , Carter Stephanie , Granchelli Joseph 
Published: August 2014

Abstract
The study of cell biology via monolayer cell culture systems is not entirely adequate for the investigation of complex inter-cell and cell to extracellular matrix interactions in vitro. Three dimensional (3D) cell culture systems better mimic more complex interactions and are extremely useful in broad applications of cell biology. In human cancer biology, a 3D culture system is better able to simulate an extracellular microenvironment to study tumor cell progression and sensitivity to anticancer agents. However, variability in forming spheres has been a persistent problem. This variability has been linked to medium composition and volume, cell density, duration in culture, and most importantly, the cellular interactions with the culture dish itself. Here, we hypothesize that a hydrophilic polymer coated surface, Thermo Scientific Nunclon Sphera, inhibits cell attachment to the culture dish by blocking binding of the extracellular matrix (ECM). First we demonstrated that the adsorption of ECM proteins (e.g. Collagen I and Fibronectin) to the hydrophilic polymer coated Nunclon Sphera surface is extremely low compared to that of the standard cell culture-treated surface, implying minimal interactions between the hydrophilic polymer and the ECM. We then examined spheroid formation on the hydrophilic polymer surface in comparison to the standard cell culture treated surface. We were able to show that unlike the standard cell culture surface, the hydrophilic polymer surface was able to support spheroid development in suspension with virtually no cell attachment by several cancer cell lines including HeLa, MCF-7, HepG2, Panc-1, Saos-2, and A549. In order to show that the hydrophilic polymer coating has no deleterious effects on cell growth, we conducted spheroid culture of a cancer stem cell line, P19.CL6, in the Nunclon Sphera dishes at relatively low cell seeding densities. Over time, the spheroids of all initial seeding densities were able to grow in volume, indicating that the hydrophilic polymer coating has no adverse effect on cell survival and proliferation. Finally, we demonstrated that the performance of Nunclon Sphera surface in supporting cancer spheroid formation is comparable to that of a similar product by a different manufacturer. Overall, the data supports that the Thermo Scientific Nunclon Sphera provides a consistent and reliable culture system for non-adherent spheroid growth, aiding in three dimensional modeling of cancer cells in vitro.
Citation Format: Cindy Neeley, Louise Gaarn, Tina Marwood, Robert Scott, Stephanie Carter, Joseph Granchelli. A novel cell culture surface supports effective formation of three dimensional cancer spheroids in suspension. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 208. doi:10.1158/1538-7445.AM2013-208