The invention discloses a frozen gel three-dimensional structural body, a preparation method and an application thereof. According to the invention, a frozen gel tissue engineering scaffold of the complex three-dimensional structure is formed, and can be used for the fields of three-dimensional cell culture, in-vitro tissue construction, tissue engineering and regenerative medicine, pathology, pharmacological study and drug testing. and the three-dimensional printing frozen gel scaffold has the characteristics of personalization, customization, high cell loading, high porosity and permeation rate, adjustable aperture size, high elastic modulus and injectable transplantation. The raw materials of the frozen gel three-dimensional structural body comprise one or a plurality of mixture of sodium alginate, gelatin, collagen, matrigel, laminin and other biomaterials and its derivative.