A low-density inoculation method for the serial subcultivation of human diploid fibroblasts: an efficient model...

Human diploid fibroblasts, TIG-1, were serially subcultivated at low inoculation densities thus maintaining the cultures in log phase throughout their in vitro lifespan. Their lifespan was shortened in calendar time without decreasing the cumulative number of population doublings when compared with the lifespan of cells serially cultivated by a standard method of 1:4 splitting. The cytokinetic and morphological changes during their lifespan are quite similar to those of control cells. The frequency of trypsinization doess not influence the cumulative number of population doublings. The "low-denstiy inoculation method" of subcultivation is efficiently and economically advantageous in studying cellular ageing in vitro.