One embodiment of the present invention provides a method and an apparatus for isolating stromal cells from biological tissue without using an enzyme, the method using the moving of stromal cells to the outside of biological tissue by inducing the spontaneous migration of the stromal cells of the biological tissue, wherein the induction of the spontaneous migration of the stromal cells is carried out in a state in which the biological tissue is attached onto an attachment member made of a material to which the biological tissue can be attached, and the induction of the spontaneous migration of the stromal cells is carried out in a culture solution where the stromal cells can survive. Therefore, problems such as the toxicity and cost of an enzyme, treatment time and risks of a heterologous virus and a problem of instability caused by using an enzyme having components extracted from the gastric fluid of animals of different species can be resolved, and, simultaneously, isolation efficiency can be improved by isolating, as stromal cells of a relatively intact undamaged natural condition, stromal cells from biological tissue without using an enzyme.