The invention discloses tissue engineering skin with a layered structure and a preparation method of the tissue engineering skin, and relates to the technical field of tissue engineering skin. The tissue engineering skin with the layered structure comprises a collagen layer, a collagen layer embedded in human skin fibroblast, a first multilayer collagen layer, a collagen layer embedded in keratinocyte and a second multilayer collagen layer. According to the tissue engineering skin disclosed by the invention, a manner that channels are innovatively generated in tissues by a temporary material such as gelatin is adopted, and human umbilical vein endothelial cells are adhered to the inner parts of the channels, so that the formed blood vessels are better fit with human body blood vessels; thetransporting and distributing rate of the formed blood vessels is increased; cells have higher viability and high multiplication rate; the printed skin with the layered structure is adopted, so thatthe interaction between cells of the formed skin structure and the interaction of the skin tissues and epimatrix are more stable, and regeneration of the skin tissue is facilitated.