Cytological changes senescing WI-38 cells: a statistical analysis

Changes in the cell, nuclear, nucleolar dry masses and areas were monitored simultaneously during in vitro senescence of WI-38 human fetal lung fibroblast cells, and the results were analyzed statistically. There was a two-step increase in all six parameters studied, following an initial decrease. The patterns of increase were strikingly similar for all six measures, i.e., an initial lag period was followed by a sharp increase, a plateau and a final, abrupt increase. In all cases, there was a strong correlation between corresponding area and dry mass measurements for each cell component. The dry mass/area ratio also increased as a function of in vitro aging and ultimately doubled in value. Weak intrapassage correlations between cell, nuclear and nucleolar dry masses indicated that there were passage to passage differences in the relative rates of dry mass increases which were not apparent over long periods of growth. The increases in all muclear and nucleolar areas were interdependent until the last doubling where they increased independently. The standard deviation for all six parameters increased as a function of in vitro senescence indicating that there was an increase in the heterogeneity of cells relative to these parameters.