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BACKGROUND: Dermal fibroblasts are widely used and demanded, and there are various isolation methods, but no comparative studies on enzyme digestion methods are reported. OBJECTIVE: To compare the cell count, morphology, migration and proliferation of human dermal fibroblasts isolated by two enzyme digestion methods. METHODS: Human dermal fibroblasts were isolated using either dispase-collagenase or trypsin, and their cell yield and viability were assessed by morphology, cell count and proliferation curve by cell counting-kit 8 assay. The ability of migration was observed by cell scratch test. RESULTS AND CONCLUSION: The fibroblasts digested with dispase-collagenase were fused at 6-7 days after inoculation, and the cells isolated by trypsin digestion were fused at 8-9 days after inoculation. Fibroblasts could be obtained by both two digestion methods. The production in the dispase-collagenase group was significantly higher than that in the trypsin group. The migration rate in the dispase-collagenase group was significantly faster than that in the trypsin group. The growth cures of the human dermal fibroblasts in the two groups revealed that the cell count was positively correlated with time, and the absorbance values of the dermal fibroblasts in the dispase-collagenase group were significantly higher than those in the trypsin group at 3, 4 and 5 days after incubation. To conclude, the cell yields, migration and proliferation of dermal fibroblasts digested with dispase-collagenase are significantly higher than those of the cells digested by trypsin, indicating that dispase-collagenase digestion results in better isolation and viability of dermal fibroblasts from the dermis. [ABSTRACT FROM AUTHOR]