The invention discloses autologous living cell soft tissue filling gel, which comprises autologous fibroblasts cultivated and amplified to 8*108, platelet-rich plasma of which the platelet concentration is 8 times that of normal blood concentration and a thrombin-calcium agent. The preparation method comprises the following steps of: amplifying the extracted fibroblasts to 8*108 in vitro; adding into the prepared platelet-rich plasma; activating with the thrombin-calcium agent; and coagulating fibrinogens in the plasma into fibrin glue with a netlike structure to obtain the living cell soft tissue filling gel, wherein culture media used in primary passage and passage of fibroblasts are prepared from a serum-free culture medium and 5 percent of platelet-rich plasma. The preparation method has the advantages that: the problems of in-vitro amplification and culturing of fibroblasts, collection and feedback, keeping of a cell healthy state in an in-vivo survival multiplication process, survival after feedback, nutrition supply of cell multiplication, space release of cell multiplication, and the like are solved, and high survival rate of cells is ensured.