The invention discloses an in vitro simulated skin model. According to an establishment method of the in vitro simulated skin model, a cell culture chamber and well plates are taken as the skeleton of the in vitro simulated skin model; the cell culture chamber is inoculated with immortalized epidermis cells, and a lower chamber well plate is inoculated with fibroblast for co-culture. The establishment method is capable of simulating the basic structure of human skin effectively; the in vitro simulated skin model can be applied in evaluation of transdermal absorption and functions in promoting fibroblast proliferation and migration of cosmetics and medicinal compounds effectively, can be used for replacing conventional animal experiments and in vitro skin tissue experiments, is capable of reducing experiment cost effectively, increasing experimental flux and efficiency, and preferably satisfying requirements on scientific research ethics with more and more attention.