Cell Survival

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: 
International Journal of Cancer. Journal International Du Cancer

Dermal fibroblasts from patients with the autosomal dominant cancer-prone disease Basal Cell Nevus Syndrome (BCNS) exhibit a serum dependence, anchorage dependence and in vitro lifespan (about 20 population doublings or less) similar to those of fibroblasts from normal age-, race- and sex-matched controls.

Author(s): 
Shimada, T.
Dowjat, W. K.
Gindhart, T. D.
Lerman, M. I.
Colburn, N. H.
Publication Title: 
In Vitro Cellular & Developmental Biology: Journal of the Tissue Culture Association

Human microvascular endothelial cells (HMVEC) from adult adipose tissue were cultured in MCDB 131 medium supplemented with 10% fetal bovine serum. Under these conditions, HMVEC from seven different donors had finite proliferative life spans ranging from 14.5 to 23.5 population doublings (PD), with a mean life span of 19 PD. Addition of 10% conditioned medium from activated human leukocyte cultures (BM Condimed) extended the life span of HMVEC to 31 to 41 PD, with a mean life span of 37 PD.

Author(s): 
Stein, G. H.
St Clair, J. A.
Publication Title: 
Mutation Research

Human diploid fibroblasts, strain MRC-5, were sequentially irradiated with 60Co gamma rays at intervals during their in vitro lifespan. The results indicate that 3 or 6 doses of 1 Gy can increase lifespan, and the same was true for cells treated with 3 doses of 3 Gy. Higher doses (5 x 3 Gy) did reduce growth potential, suggesting either that mid-late passage cells become more sensitive to radiation, or that doses beyond a given threshold reduce population lifespan by multiple cellular hits. The life extension induced by gamma rays might be due to an induced hypermethylation of DNA.

Author(s): 
Holliday, R.
Publication Title: 
Immunology Series

Table 1 summarizes the activities of hemopoietins on immature and mature basophils. IL-3, GM-CSF, and IL-5 enhanced basophil histamine release and in-vitro survival, while G-CSF, M-CSF, and IL-4 had no enhancing activities at all. In addition, IL-3 and GM-CSF induced basophil chemotaxis.

Author(s): 
Hirai, K.
Morita, Y.
Miyamoto, T.
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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