Spheroid Invasion Assay of Melanoma Cells by Hanging Drop Technique

The development of more physiologically relevant cancer models has led to the increased adoption of three-dimensional (3D) cell culture systems, such as tumor spheroids, which more accurately replicate the complex tumor microenvironment compared to traditional two-dimensional (2D) cultures. This study utilizes the hanging drop technique to generate melanoma spheroids from A375 and SK-MEL28 cell lines, including both parental and drug-resistant variants. These spheroids were embedded in Matrigel and treated with vemurafenib, cilengitide, or their combination to evaluate the effects on invasion. The combination therapy significantly reduced invasion, particularly in resistant SK-MEL28 spheroids. These findings underscore the importance of 3D spheroid models in studying drug efficacy and resistance mechanisms in cancer.