Relationship of finite proliferative lifespan, senescence, and quiescence in human cells.

Abstract

Cell hybrids were formed between human diploid fibroblasts (HDF) and carcinogen-transformed HDF to determine the relationship among: (1) finite proliferative lifespan, which we define as an age-related failure of a population to achieve one population doubling in 4 weeks; (2) arrest in a senescent state, which we define as cessation of DNA synthesis in a viable culture that is at the end of its lifespan by the above definition; and (3) arrest in a quiescent state, which we define as cessation of DNA synthesis in a young culture that is crowded or mitogen-deprived. HDF express all three of these phenotypes, which we have abbreviated FPL+, S+, and Q+, respectively. Carcinogen-transformed HDF are transformed to immortality (FPL-) and inability to achieve quiescence (Q-). They have no S phenotype because, by definition, this phenotype only exists in FPL+ cells. Fusion of FPL+, Q+, S+ HDF X FPL-, Q- carcinogen-transformed HDF produced hybrid clones that were FPL+, Q-, and S-, where the S- phenotype means that individual cells continued to synthesize DNA in cultures that had reached the end of their lifespan by our definition. These results are consistent with our hypothesis that senescent HDF and quiescent HDF may share a common mechanism for arrest in G1 phase. We have suggested that this could occur if the aging mechanism that is responsible for the FPL+ phenotype is a progressive decrease in the ability of cells to recognize or respond to mitogenic growth factors. If so, then cells would become physiologically mitogen-deprived at the end of their lifespan, which would cause them to arrest in the senescent state by the same mechanism that causes young cells to arrest in the quiescent state when they are mitogen-deprived. This hypothesis predicts that the FPL+ phenotype can be separated from the S+ phenotype--i.e., FPL+ cells can be S+ or S- --and that the Q and S phenotypes are linked--i.e., FPL+ cells are either Q+ and S+ or Q- and S-. Both these predictions are supported by the present data.