A quantitative analysis of the aging of human glial cells in culture.

The kinetics of aging of normal human diploid brain cells in culture have been determined using the miniclone technique in which cells are cloned in the presence of a large number of other cells. The miniclone technique records the behaviour of every viable cell in the sample, not merely those cells capable of forming visible clones. This technique permits the direct measurement of the reproductive potential of individual cells growing in bulk culture and of the dispersion of the sizes of colonies generated by dividing cells. The fraction of cells that are able to divide declines smoothly and continuously from the beginning of in vitro cultures of human glial cells. There is a broad distribution of colony sizes; even at the earliest passages there are significant numbers of small colonies. With increasing age of the culture there is a shift in the distribution, so that fewer large colonies and more small colonies occur. The distribution of intermitotic times is almost identical in young and middle-aged cultures. Our data seem to exclude quite positively any description in terms of a catastrophe or any abrupt change in the population. On the contrary, the decline in reproductive potential may be described adequately either as a linear change with time, or as predicted by the mortality theory of Shall and Stein (1979), in which the single constant, gamma, describes the change in reproductive potential over the entire lifetime.