Adult human skin fibroblasts were serially cultured by means of eleven protocols differing in inoculum size, duration of culture between passage and the ability of the medium to support cell division. Each protocol was terminated only when there were too few cells for further subculturing. The fraction of the cells of an inoculum adhering to the growth surface was unaffected by serial subculturing or by differences in protocol. The final cell count at the end of a period of culture and the plating efficiency for the next culture diminished progressively with serial subculturing. Nevertheless, the computed number of cell generations per culture period of those cells which divided was unaffected by serial passaging. The total number of cell doublings accruing during an entire protocol depended only on the duration of the period of culture between successive passages which was characteristic of that protocol. The observations can be accounted for quantitatively by the following assumptions. A cell which loses its ability to divide after a given period of culture nevertheless continues to grow in size during the next period of culture. The increase in volume of cell substance during any such period is the same whether or not a cell divides. The second postulate is that the probability of a cell being able to divide at the start of a period of culture is proportional to the probability that it will not lose this ability by the following period of culture.
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