In experimental cellular biology invasion is defined as disruptive proteolytic cellular movement through three-dimensional barriers Despite intense study the complex and heterogenous mechanisms of invasion and metastasis remain imperfectly understood and several mechanisms exist in collective or individual invasion of cancer cells Thus there is a need to establish robust reproducible assays to target the process and get a detailed analysis of invasion The three-dimensional 3D spheroid invasion assay is a simple and convenient method to research cell invasion which is based on the uniform and self-assembling 3D tumor spheroids