(B-80) Reducing Cancer Cell Adhesion Using Microtextured Surfaces

Introduction::
Enzymatic cell detachment strategies for cell culture are popular, but are labor-intensive, can potentially lead to accumulation of genetic mutations, and produce large quantities of waste. Thus, there is a need for surfaces that lower cell adhesion strength while maintaining cell growth to enable enzyme-free cell culture. In this study, we investigated the use of microtexture alone to control cell adhesion.




Materials and Methods::
We developed a fast, simple, and inexpensive process for creating microtextured polystyrene surfaces, a ubiquitous biocompatible material in cell culture. This fabrication method can transform any design produced with traditional lithography into a PDMS stamp with which we can mold polystyrene and can be easily scaled for high throughput cell culture as well as create high aspect ratio micron-sized features with high precision and reproducibility. We produce three different micropost surface morphologies, varying post diameter and spacing between surfaces while controlling for post top area, and compare cell adhesion strength, morphology, and viability to a flat polystyrene control surface. We quantify cell morphology and, indirectly, adhesion strength through image segmentation and analysis of our surfaces, computing average cell area, circularity, perimeter, and post contact area. We more directly measure cell adhesion strength by varying applied shear stress in a microfluidic flow channel set up and evaluating cell coverage. Doubling time and cell viability on the microtextured surfaces are assessed via a commercial cell viability analyzer, fluorescent Ki-67 expression, and Incucyte cell death assays.




Results, Conclusions, and Discussions::
These microtextured cell culture surfaces enable decreased cell adhesion strength while maintaining high cell viability and proliferation through a simple reduction in the cell-surface contact area. Using image analysis to quantify cell morphology, we found that surface textures decreased cell area by half and led to much more elongated cell shape compared to flat surfaces. With our microfluidic shear force set-up, we successfully quantified the removal of cells from the fabricated surfaces and showed that significantly more cells were removed from the microtextured surfaces than the flat surfaces, demonstrating that our surfaces lead to decreased cell adhesion. Microtextured polystyrene serves as a promising scalable alternative to reducing cell adhesion and reliance on enzymes for cell culture.




Acknowledgements (Optional): :
This work was supported by the MIT-Takeda Program Fellowship and the Broad Institute SPARC Grant.
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