The invention relates to a serum-free fibroblast cell culture medium and a preparation method thereof, wherein the base culture medium is a DMEM culture medium or a mixture of the DMEM culture medium and a F12 culture medium, and the exogenous additives comprise hormones, a metal transporter, glutamine, a lipid concentration agent, an antioxidant, purine, saccharide and a cell growth factor. Compared with the culture medium in the prior art, the serum-free fibroblast cell culture medium of the present invention has the following advantages that: animal origin or recombinant albumin is not added so as to avoid the possibly-carried safe hidden danger and reduce the culture medium cost; the culture medium is suitable for primary culture and subculture of fibroblast cells, the good cell morphology is maintained, and the status and the function of the cultured cells are similar to the status and the function of the serum group cells; adherence factor coating is not required, the primary 24 h adherence rate is more than 60%, the passage 24 h adherence rate is more than 80%, no difference exists between the cells cultured with the culture medium of the present invention and the serum group cells; requirements of multiple passages can met, and the growth conditions of the cells after 15 passages are similar to the growth conditions of the serum group cells; and after the fibroblast cells are directly transferred into the culture of the present invention from the serum-containing culture, the cells can successfully adhere and extend without the gradual adaptation process.