The invention provides serum-free culture fluid for culture of salivary gland epidermal cells and salivary gland stem cells of mammals, which is prepared by using MCDB153HAA as basic culture medium and adding amino acid, vitamins, salts, lipid, trace elements, buffer fluid, hormone-like compounds, transgenic metalloprotein, antioxidants, seralbumin, glucide, purine, pyrimidine base substances and pH value indicators. The serum-free culture fluid has the advantages that salivary gland epidermal cells of the mammals can vigorously grow, good cell activity and physiological properties are maintained, in addition, the serum-free culture fluid is also suitable for the salivary gland epidermal cell culture, is particularly suitable for the field of study relevant to biological tissue engineering and belongs to the commercial cell culture fluid.