Polystyrene-Based Slippery Surfaces Enable the Generation and Easy Retrieval of Tumor Spheroids.

Abstract

Multicellular tumor spheroids are the most well-characterized organotypic models for cancer research. Generally, scaffold-based and scaffold-free techniques are widely used for culturing spheroids. In scaffold-free techniques, the hanging drop (HD) method is a more versatile technique, but the retrieval of three-dimensional (3D) cell spheroids in the hanging drop method is usually labor-intensive. We developed oil-coated polystyrene nanofiber-based reusable slippery surfaces for the generation and easy retrieval of 3D spheroids. The developed slippery surfaces facilitated the rolling and gliding of the cell medium drops as well as holding the hydrophilic drops for more than 72 h by the virtue of surface tension as in the hanging drop method. In this study, polystyrene nanofibers were developed by the facile technique of electrospinning and the morphological evaluation was performed by scanning electron microscopy (SEM) and cryo-FESEM. We modeled the retrieval process of 3D spheroids with the ingredients of 3D spheroid generation, such as water, cell culture media, collagen, and hyaluronic acid solution, demonstrating the faster and easy retrieval of 3D spheroids within a few seconds. We created MCF-7 spheroids as a proof of concept with a developed slippery surface. 3D spheroids were characterized for their size, homogeneity, reactive oxygen species, proliferative marker (Ki-67), and hypoxic inducing factor 1ά (HIF-1ά). These 3D tumor spheroids were further tested for evaluating the cellular toxicity of the doxorubicin drug. Hence, the proposed slippery surfaces demonstrated the potential alternative of culturing 3D tumor spheroids with an easy retrieval process with intact 3D spheroids.