A process for preparing a skin culture which comprises the following steps: a) a step in which a skin culture matrix comprising a collagen sponge, a collagen sheet or a collagen gel is prepared in a means having a projection to provide said matrix with a penetrating pore, b) a step in which a skin-derived cell is seeded and cultured on said matrix, and c) a step in which the skin culture is provided with a penetrating pore by the means having the projection before said skin culture covers a skin defect, if necessary, the steps a) and b) are simultaneously carried out; a skin culture matrix and a skin culture obtained thereby. According to the present invention, a skin culture can be prepared wherein sufficient nutrition supply to epidermal cells and/or fibroblasts cultured and proliferated and discharge of exudate excessively pooled in the wound can be smoothly performed, while contraction is inhibited, by producing a penetrating pore in the culture skin before covering the wound with the culture skin with a means which requires no pretreatment in order to prevent the flowing out and losing of cells which are caused in seeding cells on a collagen matrix.
