The Effects of Glutathione on Human Keratinocyte

Cultured human keratinocytes were grown in vitro to assess the effects of glutathione, a thiol, on human keratinocyte proliferation. Single keratinocytes were harvested from newborn (greater than 5 days old) foreskin and were grown in culture flasks such that experiments could be performed in approximately 2 to 3 weeks. In order to evaluate the effects of glutathione (GSH) on keratinocyte growth, experiments were performed to determine the growth curve, plating efficiency, and incorporation of radiolabelled thymidine. Growth studies, performed with different concentrations of glutathione, revealed that 0.065 μM was the most effective concentration when compared to control cell cultures, and that the population doubling time was decreased (15.9 hrs in the presence of GSH versus 20.6 hrs in its absence). Similarly, inclusion of GSH during cell plating and radiolabelled thymidine incorporation studies revealed a positive effect on plating efficiency ( 14 % in the presence of GSH versus 9% in its absence) and mitotic index (92% in the presence of GSH versus 84% in its absence) when compared to control nontreated cultures. The research described suggests that addition of glutathione to the culture medium may improve the production of cultured epithelium by reducing the maturation time required to produce tissues usable for the treatment of severe burns.