Epidermal Growth Factor (EGF) is an endogenous peptide that promotes the process of growth, proliferation, and cell differentiation, also used for cosmetic purposes, for example as anti-aging. However, EGF has a large molecular size that can complicate its penetration into the skin. This study aims to formulate and characterize recombinant human epidermal growth factor (rhEGF) transfersomes in emulgel preparations and to study its penetration profile into the skin. In this study, transferomes formulas were optimized, namely TF1 - TF4 with several ratios of phospholipid and surfactant. The results showed that the TF3 formula with a ratio of 80:20 was the best formula with spherical and unilamellar vesicles, Z_average of 118.6 ± 1.33 nm, best deformability properties, polydispersity index of 0.102 ± 0.011, and potential zeta of -30.9 ± 0.46 mV, so this formulation was used to encapsulate rhEGF. rhEGF transfersomes were made in three formulas, namely TF-EGF1 - TF-EGF3 with different rhEGF concentrations. The best characteristics were obtained by the formula TF-EGF1 with 0.025% rhEGF concentration, which had spherical vesicles, Z_average of 128.1 ± 0.66 nm, deformability index of 1.254 ± 0.02, polydispersity index of 0.109 ± 0.004, zeta potential of -43.1 ± 1.07 mV, and encapsulation efficiency of 97.77 ± 0.06%. The three formulas were formulated into emulgel preparations (ETF1 - ETF3) and tested in vitro with non-transfersomes rhEGF emulsions (ENTF) as a comparison. The results showed that the cumulative amount of rhEGF penetrated from emulgel transferomes was higher than that which was not encapsulated in transfersomes. The ETF1 formula showed the largest increase in flux, which was 5.56 times compared to the ENTF formula. Therefore, rhEGF transfersomal emulgel has the potential to be used as an anti-aging cosmetics.