Programmable Cell Culture Chips for Topographical Manipulation of Living Cells

The micro-morphological characteristics of biomaterial surfaces play a critical role in influencing cell proliferation, adhesion, and differentiation. However, the underlying mechanisms by which surface features modulate cellular behavior remain inadequately understood. Moreover, current surface designs intended for cell regulation tend to be overly simplistic, often failing to meet the dual requirements of high-precision fabrication and structural versatility. Here, we propose a programmable cell culture chip based on femtosecond laser maskless optical projection lithography (Fs-MOPL) technology to modulate the cell behavior. The as-fabricated chip exhibits high structural fidelity and uniformity. Surface treatment with O2 plasma followed by poly-D-lysine (PDL) coating enhances hydrophilicity, cell adhesion and growth. We have investigated the migration, adhesion, and morphological changes of 786-O cells on scaffold with varied line spacing, column diameter and hole size using immunofluorescence staining and confocal fluorescence microscopy. The cells cultured on linear array structures display elongated, oriented actin stress fibers, while column and hole array structures influence focal adhesion distribution and cellular tension. Biocompatibility characterization further confirms the chip's suitability for cell culture applications. Our findings highlight the potential of programmable cell culture chips to mimic complex in vivo microenvironments, offering a multifunctional platform for studying cell behavior and advancing biomedical research.