Abstract: The present disclosure discloses a preparation method and a recovery method of pariduval mesenchymal stem cells (PMSCs). In the preparation method, a high-glucose Dulbecco's Modified Eagle Medium (DMEM) that includes a Tryple-ethylenediaminetetraacetic acid (EDTA) enzyme of 40% to 60% in volume concentration and collagenase type II of 8 mg/ml to 12 mg/ml is used as a tissue digestion solution to digest tissue blocks, which facilitates PMSCs to climb out of the tissue blocks and grow adherently; and a serum-free DMEM is adopted as a selective medium to terminate the digestion and resuspend PMSCs, which helps to improve a purity of PMSCs, accelerate the growth of PMSCs, and achieve the rapid expansion of PMSCs in vitro.