Aim To discuss a set of system to culture vascular smooth muscle cell(VSMC),vascular adventitial fibroblast(VAF) and vascular endothelial cell(VEC) from murine aorta,which are simple,reliable and easy to replicate. Methods The primary culture of VSMC,VAF and VEC used the tissue adherent method,trypsin digestion for passage transfer. The cells were purified by differential adherent and natural growth purification. Myofibroblast(MF) was transformed from VAF by growth factor-β1(TGF-β1) induced. Phase contrast microscope and immunocytochemistry staining were used to identify the morphological and the immunological characteristics,respectively. Results The tissues and cells activity were excellent. The growth cycle of primary VSMC/VAF and VEC were 10~12 days and 12~14 days,respectively. And passage cycle was 7~10 days. After being purified and subculture,the cells purity achieved 95%~100%. The growth characteristics of VSMC assumed the model "peak and valley",and with anti α-SMA(+) /Vimentin(-);the VEC assumed "pebble-like" appearance,and anti CD31(+)/α-SMA(-);the staining of VAF were Vimentin(+)/α-SMA(-),while the induction MF were Vimentin(+)/α-SMA(+). The primary cells were successfully passaged more than 10 generations,without growth vigor decreasing. Conclusion We have established a set of system methods to culture the primary VEC,VSMC,VAF and MF of rat aorta origin,the advantages of our approach are simple,reliable and easy to replicate,while the cells have high purity and excellent biological activity. This method is still suitable for culturing the ingredient cell of mouse aorta vessel wall.