Pontén J, Stein W D, Shall S
J Cell Physiol. 1983 Dec;117(3):342-52. doi: 10.1002/jcp.1041170309.
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.
利用微克隆技术测定了体外培养的正常人二倍体脑细胞的衰老动力学。在微克隆技术中，细胞是在大量其他细胞存在的情况下进行克隆的。微克隆技术记录样本中每个活细胞的行为，而不仅仅是那些能够形成可见克隆的细胞。该技术允许直接测量在批量培养中生长的单个细胞的繁殖潜力以及分裂细胞产生的集落大小的离散度。从人神经胶质细胞体外培养开始，能够分裂的细胞比例就平稳且持续下降。集落大小分布广泛；即使在最早传代时，也有大量小集落。随着培养时间的增加，分布发生变化，大集落减少，小集落增多。年轻和中年培养物中间期时间的分布几乎相同。我们的数据似乎可以肯定地排除用灾难或群体中的任何突然变化来描述的情况。相反，繁殖潜力的下降可以充分地描述为随时间的线性变化，或者如沙尔和斯坦因（1979年）的死亡率理论所预测的那样，其中单一常数γ描述了整个生命周期中繁殖潜力的变化。
