The invention relates to a method for establishing and culturing Tianzhu Yak'ear marginal fibroblast cell line, utilizing the fast tissue wall pasting method to improve culture medium, and finally obtaining high-yield and high-purity Tianzhu Yak'ear marginal fibroblast cell line whose conservation number is CGMCC No.1881, belonging to the field of cell biology. And the frozen cells have stable quality, suitable to large scale culture. And the Tianzhu Yak'ear marginal fibroblast cell line can provide large numbers of high quality materials for research on science of living systems in gene engineering, cell engineering, immunity and molecular biology; can act as donor cells for animal's somatic cell cloning and breeding; and can enrich local variety improvement and storage of local excellent variety in agriculture.