Abstract: Objective　To explore a simple and efficient method for the isolation of fibroblasts, and to evaluate the outcomes of fibroblasts isolation using enzyme digestion without filtration from keloids. Methods　Keloid tissues were divided into three parts with the same volume, and cells were isolated using enzyme digestion without filtration, traditional enzyme digestion and tissue cultivation respectively. Cell growth was observed and the time required for cells to coat 80% of the bottom of the cell culture flasks was recorded. In addition, the cell proliferation and the function of collagen secretion in each group were detected and compared. Results　In the group of enzyme digestion without filtration, there were small bud-like adherent cells swimming out from the tissue debris in only 2-3 days after plating and the single adherent cells grew well. The time required for cells to coat 80% of the bottom of the flasks in this group was 3.25±0.89 days, less than that in the traditional enzyme digestion group, and 9.50±1.64 days less than that in the tissue cultivation group (P<0.01). Contamination was not found in all groups. There were no significant differences in the growth rate of 24-hour cell proliferation activity and the hydroxyproline content among the three groups (P>0.05). Conclusion　Enzyme digestion without filtration contains fewer steps and is easy to operate, which will not increase the rate of contamination or influence the cell viability or the function of collagen secretion. It is an efficient method for fibroblasts extraction and worth spreading.

Key words: Scar, 　Keloid, 　Fibroblasts, 　Cell isolation, 　Enzyme digestion