Abstract: In order to study dedifferentiation and cell orientation in the body of human adult cell, adult human fibroblast lines that stably expressed exterior gene were established. Adult male foreskin subcutaneous tissues apart from epidermis and hypodermis were taken by surgery operation and human foreskin fibroblasts were isolated and cultured, then enhanced green fluorescence protein (EGFP) in human fibroblasts were expressed by lipofectamine and lentiviral vectors, respectively. Results showed that human fibroblasts had shuttle-like image and stable growth characteristics. EGFP was unsteadily expressed in human fibroblasts, positive cells in total fibroblasts were very low, human fibroblasts became thinner and had slower growth characteristic after transfected by lipofectamine. In contrast, EGFP was steadily expressed in human fibroblasts, positive cells in total fibroblasts was very high, and fibroblasts had rapid proliferation and stable growth characteristics that were not affected by continous cell passage and cell frozen-thawn after transfected by lentiviral vectors. In addition, EGFP positive fibroblasts in total cells were 99.85% and homogeneous fibroblasts in total cells were 76.05% by flowcytometer assay. It is showed that expression of EGFP gene in human fibroblasts mediated by lentiviral vector was high efficient and stable.