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In this work, electrospinnable and water stable soyprotein was extracted by using a reducing agent in mild alkaline condition, and novel 3D zein and 3D pure soyprotein electrospun scaffolds with three-dimensionally and randomly oriented fibers and large interconnected pores were successfully fabricated by reducing surface resistivity of materials. This unique structure is different from most electrospun scaffolds with fibers oriented mainly in one direction. The structure of novel 3D scaffolds could more closely mimic the 3D randomly oriented fibrous architectures in many native extracellular matrixes (ECM). Confocal laser scanning microscope shows that instead of becoming flattened cells when cultured in conventional electrospun scaffolds, the cells cultured on novel 3D scaffolds could develop into stereoscopic topographies, which highly simulated in vivo 3D cellular morphologies and are believed to be of vital importance for cells to function and differentiate appropriately. In vitro cell attachment, proliferation and differentiation study indicated that the 3D fibrous scaffold could better support the attachment and proliferation of NIH 3T3 mouse fibroblast cells, and could better support ADMSC for proliferation and adipogenic differentiation. One mechanism of this fabrication process has also been proposed and shown that the rapid delivery of electrons on the fibers was the crucial factor for formation of 3D architectures. The novel dissolution method could be applied to a number of water stable proteins that contains large amount of intermolecular and intramolecular disulfide bond crosslinkages, and 3D electrospinning method could be applied to many other proteins and materials.