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One of the major obstacles in developing drugs for diabetes is the lack of in vitro models that can capture more features of the dynamic insulin secretion. Here, we engineered an insulin secretion model by cell-assembly technique which can assemble cells into designated places to form complex three-dimensional structures. Adipose-derived stromal cells were assembled and induced differentiation into adipocytes; pancreatic islets were then assembled at designated stations. A perfusion system was used for a time-dependent estimation of insulin response to glucose stimulation. After long-term exposure to high glucose, the insulin secretion peak value of β-cell in the 3D system decreased and delayed compared with the 2D culture system. These results suggested that adipocytes in 3-D structure help β-cells capture more pathologic features of diabetes. Drugs, Nateglinide and rosiglitazone, known to have effects on diabetes showed accordant effect in the system. This 3D culture model provides an excellent system to study the insulin secretion kinetics of the β-cells. This study also demonstrates that the 3D culture system can provide a model for diabetes drug discovery.