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The establishment of primary keloid fibroblast culture has always been a fundamental measure for studying mechanisms of keloid disease. The quality of the primary cell culture can directly affect the results of further experiments. T h s study was performed to investigate the optimal growth conditions, including the optimal storage time and collagenase treatment time, for in vitro cell culture models and the suitable methods for epidermis-dermis separation in different tissues. Keloid tissues, keloid-surro unding tissues, and normal skin tissues were collected from patients, for primary fibroblast culture. Two methods, tissue explant and col genase digestion, were deployed and compared. Expression levels of the keloid-related genes α-SMA, Col1, and Col3 were assessed in cells cultured using both methods, to verify the qualities of the primary cells. A comparative analysis was conducted between the two met ods and among the three different tissues used. Bacterial and lipid contamination was immediately minimized after the samples were processed. Different methods of epidermis removal and different durations of collagenase digestion were required in different tissues to g enerate optimal results. Real-time PCR results showed that the mRNA expression levels of keloid-related genes in cultured fibroblasts c or elated to their in vivo expression profile, as previously reported in other studies. The results of this study have revealed several k ey points in the culture of primary keloid fibroblasts and demonstrated the correlation in gene expression between in vivo keloid fibroblasts an d in vi ro primary keloid fibroblasts.

international journal of morphology

Comparison of Primary Keloid Fibroblast Cultivation Methods and the Characteristics of Fibroblasts Cultured from Keloids, Keloid-surrounding Tissues, and Normal Skin Tissues