Expression of TGF-β۳ in Isolated Fibroblasts from Foreskin

Background: The multifunctional transforming growth factor beta (TGF-β) is a glycoprotein that exists in three isoforms. TGF-β۳ expression increases in fetal wound healing and reduces fibronectin and collagen I and III deposition, and also improves the architecture of the neodermis which is a combination of blood vessels and connective tissue during wound healing. Fibroblasts are key cells in the wound healing process. TGF-β۳ plays a critical role in scar-free wound healing and fibroblast actions in the wound healing process. The aim of this study was to express the TGF-β۳ gene (tgf-b۳) in human foreskin fibroblasts (HFF’s). Methods: We obtained HFF’s from a newborn and a primary fibroblast culture was prepared. The cells were transfected with TGF-β۳-pCMV۶-XL۵ plasmid DNA by both lipofection and electroporation. Expression of TGF-β۳ was measured by enzyme-linked immunosorbent assay (ELISA). Results: The highest TGF-β۳ expression (۸.۳-fold greater than control) was obtained by lipofection after ۷۲ hours using ۳ µl of transfection reagent. Expression was ۱.۴-fold greater than control by electroporation. Conclusions: In this study, we successfully increased TGF-β۳ expression in primary fibroblast cells. In the future, grafting these transfected fibroblasts onto wounds can help the healing process without scarring.

Keywords:

Fibroblasts , Gene expression , TGF-β۳