The study on 4D culture system of squamous cell carcinoma of tongue.

Traditional cell culture does not accurately simulate the cell microenvironment and demonstrate the specificity of individual cell growth. In this study, we created a 4D cell culture model. It is a precision instrument consisting of an electromagnet, a force transducer, and a cantilever bracket. A petri dish is placed above the magnet, gel beads encapsulated with magnetic nanoparticles and tongue cancer cells are placed in the petri dish. In this model, a magnetic force is generated on the magnetic nanoparticles in the culture medium to drive gel movement when the magnet is energized, and the gel exerts an external force on the cells. It can mimic the microenvironment inside the tongue squamous cell carcinoma cell CAL-27 when the tongue is moving. Electron microscopy and rheological analysis experiments were performed on hydrogels to characterize the alginate. Calcein-AM/PI staining was conducted to verify the biosafety of the hydrogel culture system. Four experimental groups were set up in this study. On this basis, we confirmed that proliferation of tongue squamous cell CAL-27 significantly increased after 5 days compared to cells cultured without mechanical stimulation by MTT. The cell diameter of different groups was measured on captured photographs, and it was concluded that the cell diameter in the dynamic culture environment was larger than that of the hydrogel culture alone in the same period. Furthermore, the cell morphology of tongue squamous cell carcinoma was better. In flow cytometry experiments, there were fewer cells in the G0/G1 phase while the proportion of cells in the G2/M phase was increased following mechanical stimulation. The changes in RNA levels between groups after 21 days of culture were detected using RT-PCR. This device, mimicking the microenvironment of tongue cancer cells in vivo, can enable better visualization of the cell growth in vivo. Therefore, this study provides a reliable basis for subsequent tongue biopsy, research on the pathogenesis of tongue cancer, and drug treatment.&#xD.