Treating chronic wound is challenging, especially in unhealthy patients. Recently, the use of allogenic keratinocyte grafting has been shown to improve wound healing. We report seven cases with recalcitrant wounds which were treated successfully by using a biological dressing material derived from cultured human keratinocytes. The skin wounds in these cases had been resistant to the various treatments including occlusive dressing, polydeoxyribonucleotide injection, and platelet-rich plasma therapy. The biological dressing material derived from cultured human keratinocytes produced noticeable improvements in the recalcitrant wounds without any complications. We think that the biological dressing played an important role in wound healing. Therefore we suggest a biological dressing material derived from cultured human keratinocytes can be considered as a treatment option for ulcerative wounds.