The choice of artificial extracellular matrix or scaffold that could offer skin stem cells optimal environment for in vitro growth and differentiation is particularly important. Scaffolds for skin engineering must meet three basic criteria: patient safety, clinical efficacy and convenience of use. The aim of this study was to create in vitro fibrin based substitute with epidermal and dermal component and to evaluate treatment of deep partial and full thickness burns. In this study human skin samples and skin cells from surgical surplus tissue were used. Under highly controlled conditions fibroblasts and keratinocytes were cultured. From commercial fibrin glue kits, fibrin scaffolds were aseptically prepared. For viability tests, scanning electron microscope (SEM) and immunocytochemistry analysis of cells cultured on fibrin scaffold were performed. After hydrosurgical preparation of deep burn necrotic tissue, wound bed was prepared for caltar and skin equivalents. Progress of healing was documented using a drawing chart and photos. In epithelial cultures on feeder layer, keratinocytes had characteristic polygonal morphology, while dermal fibroblasts showed bipolar spindle morphology. SEM images showed good attachment and colony spreading of keratinocytes and fibroblasts on fibrin scaffold. Immunofluorescent staining of keratinocyte cultures on fibrin scaffold showed expressions of CK19 (epithelial stem cells marker), involucrin, and marker of differentiated keratinocytes. Clinical results have clearly shown that appearance of the skin did not differ significantly from areas of transplanted skin using standard techniques. In conclusion, using fibrin-cultured autografts on massive full-thickness burn would result in good healing up to 3 years of follow-up.