The cancer cells in brain tumors interact with their microenvironment, which includes stromal cells, the extracellular matrix (ECM), and the physical properties of tissues. The reciprocal interaction between cancer cells and the surrounding microenvironment regulates the biological behavior of cancer cells. To improve our understanding of the progression of brain tumors, it is useful to construct physiologically relevant brain tumor models. Consequently, versatile in vitro tumor models ranging from simplistic two-dimensional (2D) cultures to three-dimensional (3D) cultures have been developed to mimic the microenvironments of the brain. In this dissertation, we developed that 1) characterization of tissue-derived extracellular matrix (tdECM), 2) reconstruction of patient brain tissue-derived extracellular matrix (pdECM)-based in vitro model, and 3) application of our in vitro 3D brain tumor model.