Glutathione content of cultured human fibroblasts during in vitro ageing

The glutathione level of cultured human fibroblasts was determined with a micromodification of a spectrophotometric glutathione cycling method. There was a slight increase in reduced glutathione (GSH) content during in vitro ageing of normal human fibroblasts. Fibroblasts from patients with Werner's syndrome or ceroid lipofuscinosis (Spielmeyer-Vogt syndrome) and healthy individuals exhibited similar patterns of GSH levels during in vitro ageing. The GSH content of non-proliferating confluent cultures of normal fibroblasts and of proliferating normal fibroblasts was identical. Moreover, autofluorescent "aged" cells isolated by cell sorting did not differ in GSH content from the non-autofluorescent cells in the same culture. It was concluded that the GSH content does not play a role in in vitro ageing, nor in the accumulation of autofluorescent material in human skin fibroblasts.