We have established culture conditions for cells from normal limb, early limb regenerate (blastema), heart, and liver of the newt Notophthalmus viridescens. Whereas heart and liver cells had a relatively short life in culture, limb cells have shown no sign of senescence over more than 1 year in culture. Cultured cells from all these tissues express to differing extents the regeneration-associated antigen 22/18. The antigen is intracellular and filamentous, and its expression appears to be regulated by culture density. Furthermore, 22/18 antigen is turned off in limb and blastemal cultures following differentiation into muscle, as also occurs in vivo.