An interplay of matrix stiffness, dimensionality and adhesivity on cellular behavior.

Cell-based assays are essentialin vitrotools for understanding basic cell biology, pathophysiology of diseases and mechanism of drug actions most cancer studies have utilized two-dimensional (2D) cell culture methods, which have their shortcomings including lack of cell- extracellular matrix interactions and three-dimensional (3D) geometry, and inaccurate representation of cell polarity. Hence, 3D matrices are being increasingly used to study the effect of 3D niche on cell behavior. Till date, very few systematic studies have been done to show comparison of cell behavior when seeded on the surface and encapsulated inside the matrix. In this study, we fabricated poly(ethylene glycol) (PEG) and gelatin-based matrices using UV mediated photo-polymerization to establish 2D and 3D cell culture methods using breast cancer MDA-MB-231 cells. We have found that the adhesion and spreading of cells on the gel surface is different from that when embedded in gels. The stiffness of poly (ethylene glycol) diacrylate (PEGDA)-gelatin methacryloyl (GelMA) hydrogels with lower concentration of GelMA is lower than that with higher GelMA; further, those with higher overall concentration of polymers (>5%) retain their mechanical integrity and do not degrade even after 7 d. Physical characterization of these matrices demonstrate their optimal pore size, mechanical stiffness and degradation, which are further tunable for tissue engineering, regenerative medicine, drug delivery and cancer studies. Additionally, these semi-synthetic PEGDA-GelMA matrices are transparent in nature, thereby, allowing easy imaging of cells in 3D. The system developed here can be used for short and long term cell culture and can be potentially explored for cell migration and metastasis studies.