〔Abstract〕 Objective To isolate, culture and identify adult mouse dorsal keratinocyte(KC) in vitro and to compare of the proliferation and differentiation capabilities of four adult mouse dorsal skin KC. Methods Four kinds of adult mouse dorsal skin were taken, the subcutaneous adipose tissue was peeled, and KC was obtained by 0.25% trypsin digestion; KC were identified using immunofluorescence method,cell growth status was observed under microscopy,different seed plate times and densities were designed,and cell proliferation viability was detected by CCK-8,Ed U and high connotation method. Four adult mouse KC differentiation was detected by Western blot. Results Trypsin digestion was used to extract four strains of adult mouse back skin KC,and the Kunming( KM)mice had the lowest number of KC and C57BL/6 to extract the highest number of cells for the same area of skin tissue. BALB/c,KM,and nude mouse had higher plate-laying efficiency at a density of 8 × 103/well,respectively,and C57BL/6 had higher plate-laying efficiency at a density of 1. 6 × 104/well. Among the four strains of mice,the primary KC of BALB/c mice was weaker than that of C57BL/6 mice,and there was no statistical difference in the degree of differentiation. Conclusion The proliferation and differentiation capacity of four adult mouse primary KC on the dorsal skin is successfully isolated and compared by a simple and easy method,which provided a certain experimental basis for the strain selection of mice with skin-related diseases.