Changes in DNA alkali-sensitive sites during senescence and establishment of fibroblasts in vitro

DNA molecular weight was studied in human embryonic and mouse newborn lung fibroblasts in vitro at different passages of the culture using alkaline and neutral sucrose gradient techniques. Reduction of molecular weight of single-stranded DNA due to alkaline-sensitive sites appeared spontaneously during the growth decline of the mouse cells. These changes disappeared when the mouse fibroblasts became a permanent cell line. At the end of phase II of the human fibroblasts, the molecular weight of single-stranded DNA also decreased, followed by the restitution of some high molecular weight DNA in the ultimate passages. When treated with 1 mM caffeine, the mouse fibroblasts during growth crisis did not survive, while cells of the established line resisted. Thus it might be possible that a DNA repair process was involved in the recovery of the mouse fibroblasts. Furthermore, results favor the hypothesis that the cells that become established are not present in the primary culture but originate in vitro.