Abstract: A method of making a tissue reference control block is disclosed wherein cancer cells are selected with a known positivity or negativity for a specific marker and cancer cells are grown to a certain phase. The cancer cells are screened for an expression of the specific marker and the screened cancer cells are mixed with stromal cells and co-cultured in a bio-reactor bag. A spheroid grown from the co-cultured mixture of cells is harvested, fixed, and embedded in a core of a paraffin block.

Abstract: The disclosed embodiments include methods to form STC and STMC for use in determining presence of cancer, and methods to detect presence of cancer. In one embodiment, A portion of a STC is stained. The STC includes normal cells and cancer cells of a type of cancer co-cultured based on at least one cell culturing factors. The at least one co-culture factors includes a type of the cancer cells being cultured, a ratio of the cancer cells to the normal cells being co-cultured, seeding density of the normal cells and the cancer cells being co-cultured, a type of cell growth supplement used to facilitate culturing the cells, and a concentration of the cell growth supplement used to facilitate co-culturing the cells. The stained portion is observed to determine presence of one or more biomarker types that indicate presence of cancer.