Cell Line

Publication Title: 
Proceedings of the National Academy of Sciences of the United States of America

Inclusion of vitamin E (DL-alpha-tocopherol) in the culture medium for human diploid cells greatly prolongs their in vitro lifespan. The addition of 100 mug of DL-alpha-tocopherol per ml of medium has allowed us to culture WI-38 cells for more than 100 population doublings to date. (These cells normally have an in vitro lifespan of 50 +/- 10 population doublings.) Cells at the 100th population doubling have a normal diploid karyotype, appear to behave in all other respects like young WI-38 cells, and are still actively dividing.

Author(s): 
Packer, L.
Smith, J. R.
Publication Title: 
Advances in Experimental Medicine and Biology

It has been shown that human diploid cells from various donor ages can be arrested in an essentially nonmitotic state by reducing the serum concentration of the incubation medium from 10 to 0.5 percent. Cells incubated at this serum level maintained the population distribution that was present when the cells reached confluency. The population, which has 90 percent of the cells in the G1 phase of the division cycle, was not static and exhibited a low level of mitotic activity with prolonged interdivision times.

Author(s): 
Dell'Orco, R. T.
Publication Title: 
Proceedings of the National Academy of Sciences of the United States of America

Previously we reported [Packer, L. & Smith, J.R. (1974) Proc. Natl. Acad. Sci. USA 71, 4763-4767] that the lifespan of WI-38 human diploid fibroblasts in vitro was significantly increased by continuously growing the cell cultures in the presence of vitamin E(dl-alpha-tocopherol), but in 19 subsequent subcultivation series we were unable to reproduce these findings. While vitamin E is incorporated into the cells and is able to act effectively as an antioxidant, apparantly is intracellular antioxidant properties alone do not routinely result in an increase of cell lifespan.

Author(s): 
Packer, L.
Smith, J. R.
Publication Title: 
Journal of Cellular Physiology

Various concentrations of oxygen were used to determine the optimum culture medium PO2 for survival and proliferation of attached human and mouse fibroblasts grown from different inoculum sizes. When T-15 flasks were seeded with less than or equal to 2 X 10(4) cells (less than or equal to 1.3 X 10(3) cells/cm2), the highest plating efficiencies and cell yields were obtained with a culture medium PO2 of 40-60 mm Hg.

Author(s): 
Taylor, W. G.
Camalier, R. F.
Sanford, K. K.
Publication Title: 
Experimental Cell Research

Normal human diploid cells, TIG-1, ceased to proliferate at about the 62 population doubling level (PDL). Transformed clones isolated from TIG-1 cells infected with wtSV40 and those with tsA900 SV40 cultured at 34 degrees C were subcultured up to about 80 PDL. When the culture temperature of tsA SV40-transformed cells was shifted from 34 to 39.5 degrees C at 51 PDL, the growth curve of these transformed cells changed to that of normal young cells.

Author(s): 
Ide, T.
Tsuji, Y.
Nakashima, T.
Ishibashi, S.
Publication Title: 
Journal of Cellular Biochemistry

Tumor-promoting phorbol esters, like growth factors, elicit pleiotropic responses involving biochemical pathways that lead to different biological responses. Genetic variant cell lines that are resistant to mitogenic, differentiation, or transformation responses to tumor promoters have been valuable tools for understanding the molecular bases of these responses.

Author(s): 
Colburn, N. H.
Smith, B. M.
Publication Title: 
Experimental Cell Research

Lymphocytes have a finite and predictable proliferative life span in culture similar to that observed in fibroblasts. In general, the senescence of human fibroblasts is inevitable and irreversible, but their proliferative life span can be extended by certain DNA tumor virus oncogenes, such as the large T antigen of the SV40 virus. Here, we show that human T lymphocytes (HTL) can be stably transfected with SV40 large T and that expression of T antigen extended the life span of T cell cultures.

Author(s): 
Ryan, Q. C.
Goonewardene, I. M.
Murasko, D. M.
Publication Title: 
Experimental Cell Research

Cellular senescence is a state of irreversible cell cycle arrest in which normal cells at the end of their lifespan fail to enter into DNA synthesis upon serum or growth factor stimulation. We examined whether proteins required for G1/S cell cycle progression were irreversibly down-regulated in senescent human fibroblasts. Both the 44- and 42-kDa forms of the MAP-kinase protein were expressed at similar levels in young and senescent cells.

Author(s): 
Afshari, C. A.
Vojta, P. J.
Annab, L. A.
Futreal, P. A.
Willard, T. B.
Barrett, J. C.
Publication Title: 
Critical Reviews in Oncogenesis

For several decades simian virus 40 (SV40) early region genes have been used as a means of generating immortalized human cell lines; however, the molecular mechanisms of this process have begun to be understood only recently. SV40-induced immortalization proceeds via two phases. In the first phase ("lifespan extension"), cells continue proliferating for a limited number of population doublings beyond the point at which normal cells undergo senescence.

Author(s): 
Bryan, T. M.
Reddel, R. R.
Publication Title: 
Oncogene

Normal human breast epithelial cells were transfected with expression vectors containing the p53 gene mutated at either codon 143, 175, 248 or 273, or by infection with a recombinant retroviral vector containing the p53 gene mutated at codons 143, 175, 248, or 273. The breast epithelial cells were monitored for extension of in vitro lifespan and immortalization. Expression of some, but not all, p53 mutants resulted in an extension of in vitro lifespan.

Author(s): 
Gollahon, L. S.
Shay, J. W.

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