This protocol describes the isolation and primary culture of stem cells from rabbit adipose tissue. Adipose-derived stem cells (ADSCs) are a valuable source of multipotent cells with potential applications in regenerative medicine, tissue engineering, and cell therapy. The procedure involves harvesting adipose tissue, enzymatic digestion using the HyQTase enzyme, and subsequent isolation of the stem cells through centrifugation and washing steps. Following isolation, cell viability is assessed using Trypan Blue staining, and the stem cells are cultured in a medium supplemented with fetal bovine serum and antibiotics. This method provides a reliable approach for obtaining and cultivating ADSCs for downstream applications in research and therapy.