Abstract
Human diploid cells (WI-38) were serially subcultivated at partial pressures of oxygen (Po2) ranging from 5.6 mm Hg to 608 mm Hg. At a Po2 of 5.6 mm Hg, the number of doublings to phase out was less than that of control cells at a Po2 of 137 mm Hg. Cultures grown at Po2's of 24, 49, or 137 mm Hg grew at the same rate and phased out after a similar number of population doublings. Population lifespan was markedly shortened by chronic exposure to elevated Po2's, a phenomenon that was, in part, reversible. d-1-alpha-Tocopherol (10 microgram/ml or 100 microgram/ml) homogenized into the medium at each weekly subcultivation did not extend the lifespan of cells at reduced, ambient, or elevated oxygen tensions. These results indicate that neither oxygen toxicity nor free radical reactions play a significant role in limiting the lifespan of WI-38 cells grown in vitro under ambient oxygen tensions (Po2 137 mm Hg).
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- BURK D., SCHADE A. L. On respiratory impairment in cancer cells. Science. 1956 Aug 10;124(3215):270–272. [PubMed] [Google Scholar]
- Balin A. K., Goodman B. P., Rasmussen H., Cristofalo V. J. The effect of oxygen tension on the growth and metabolism of WI-38 cells. J Cell Physiol. 1976 Oct;89(2):235–249. doi: 10.1002/jcp.1040890207. [DOI] [PubMed] [Google Scholar]
- Balin A. K., Goodman D. B., Rasmussen H., Cristofalo V. J. Atmospheric stability in cell culture vessels. In Vitro. 1976 Oct;12(10):687–692. doi: 10.1007/BF02797472. [DOI] [PubMed] [Google Scholar]
- CHANCE B., COHEN P., JOBSIS F., SCHOENER B. Intracellular oxidation-reduction states in vivo. Science. 1962 Aug 17;137(3529):499–508. doi: 10.1126/science.137.3529.499. [DOI] [PubMed] [Google Scholar]
- Chio K. S., Reiss U., Fletcher B., Tappel A. L. Peroxidation of subcellular organelles: formation of lipofuscinlike fluorescent pigments. Science. 1969 Dec 19;166(3912):1535–1536. doi: 10.1126/science.166.3912.1535. [DOI] [PubMed] [Google Scholar]
- Comfort A. The position of aging studies. Mech Ageing Dev. 1974 Mar;3(1):1–31. doi: 10.1016/0047-6374(74)90002-5. [DOI] [PubMed] [Google Scholar]
- Comfort A., Youhotsky-Gore I., Pathmanathan K. Effect of ethoxyquin on the longevity of C3H mice. Nature. 1971 Jan 22;229(5282):254–255. doi: 10.1038/229254a0. [DOI] [PubMed] [Google Scholar]
- Corwin L. M., Humphrey L. P. Vitamin E: substrate-dependent growth effect on cells in culture. Proc Soc Exp Biol Med. 1972 Nov;141(2):609–612. doi: 10.3181/00379727-141-36833. [DOI] [PubMed] [Google Scholar]
- Cristofalo V. J., Sharf B. B. Cellular senescence and DNA synthesis. Thymidine incorporation as a measure of population age in human diploid cells. Exp Cell Res. 1973 Feb;76(2):419–427. doi: 10.1016/0014-4827(73)90394-7. [DOI] [PubMed] [Google Scholar]
- Daniel C. W., De Ome K. B., Young J. T., Blair P. B., Faulkin L. J., Jr The in vivo life span of normal and preneoplastic mouse mammary glands: a serial transplantation study. Proc Natl Acad Sci U S A. 1968 Sep;61(1):53–60. doi: 10.1073/pnas.61.1.53. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Daniel C. W., Young L. J. Influence of cell division on an aging process. Life span of mouse mammary epithelium during serial propagation in vivo. Exp Cell Res. 1971 Mar;65(1):27–32. doi: 10.1016/s0014-4827(71)80046-0. [DOI] [PubMed] [Google Scholar]
- Deamer D. W., Gonzales J. Autofluorescent structures in cultured WI-38 cells. Arch Biochem Biophys. 1974 Nov;165(1):421–426. doi: 10.1016/0003-9861(74)90181-7. [DOI] [PubMed] [Google Scholar]
- EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
- EAGLE H. The minimum vitamin requirements of the L and HeLa cells in tissue culture, the production of specific vitamin deficiencies, and their cure. J Exp Med. 1955 Nov 1;102(5):595–600. doi: 10.1084/jem.102.5.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
- GOLDBLATT H., CAMERON G. Induced malignancy in cells from rat myocardium subjected to intermittent anaerobiosis during long propagation in vitro. J Exp Med. 1953 Apr;97(4):525–552. doi: 10.1084/jem.97.4.525. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grove G. L., Mitchell R. B. DNA microdensitometry as a measure of cycling-non-cycling activity in aged human diploid cells in culture. Mech Ageing Dev. 1974 Sep-Oct;3(3-4):235–240. doi: 10.1016/0047-6374(74)90019-0. [DOI] [PubMed] [Google Scholar]
- HARMAN D. Aging: a theory based on free radical and radiation chemistry. J Gerontol. 1956 Jul;11(3):298–300. doi: 10.1093/geronj/11.3.298. [DOI] [PubMed] [Google Scholar]
- HARMAN D. Role of free radicals in mutation, cancer, aging, and the maintenance of life. Radiat Res. 1962 May;16:753–763. [PubMed] [Google Scholar]
- HAYFLICK L., MOORHEAD P. S. The serial cultivation of human diploid cell strains. Exp Cell Res. 1961 Dec;25:585–621. doi: 10.1016/0014-4827(61)90192-6. [DOI] [PubMed] [Google Scholar]
- HAYFLICK L. THE LIMITED IN VITRO LIFETIME OF HUMAN DIPLOID CELL STRAINS. Exp Cell Res. 1965 Mar;37:614–636. doi: 10.1016/0014-4827(65)90211-9. [DOI] [PubMed] [Google Scholar]
- HENDLEY D. D., MILDVAN A. S., REPORTER M. C., STREHLER B. L. THE PROPERTIES OF ISOLATED HUMAN CARDIAC AGE PIGMENT. II. CHEMICAL AND ENZYMATIC PROPERTIES. J Gerontol. 1963 Jul;18:250–259. doi: 10.1093/geronj/18.3.250. [DOI] [PubMed] [Google Scholar]
- Harman D. Prolongation of life: role of free radical reactions in aging. J Am Geriatr Soc. 1969 Aug;17(8):721–735. doi: 10.1111/j.1532-5415.1969.tb02286.x. [DOI] [PubMed] [Google Scholar]
- KROHN P. L. Review lectures on senescence. II. Heterochronic transplantation in the study of ageing. Proc R Soc Lond B Biol Sci. 1962 Dec 18;157:128–147. doi: 10.1098/rspb.1962.0066. [DOI] [PubMed] [Google Scholar]
- Kilburn D. G., Lilly M. D., Self D. A., Webb F. C. The effect of dissolved oxygen partial pressure on the growth and carbohydrate metabolism of mouse LS cells. J Cell Sci. 1969 Jan;4(1):25–37. doi: 10.1242/jcs.4.1.25. [DOI] [PubMed] [Google Scholar]
- Le Guilly Y., Simon M., Lenoir P., Bourel M. Long-term culture of human adult liver cells: morphological changes related to in vitro senescence and effect of donor's age on growth potential. Gerontologia. 1973;19(5):303–313. doi: 10.1159/000211984. [DOI] [PubMed] [Google Scholar]
- Levine E. M. Mycoplasma contamination of animal cell cultures: a simple, rapid detection method. Exp Cell Res. 1972 Sep;74(1):99–109. doi: 10.1016/0014-4827(72)90484-3. [DOI] [PubMed] [Google Scholar]
- Lipetz J., Cristofalo V. J. Ultrastructural changes accompanying the aging of human diploid cells in culture. J Ultrastruct Res. 1972 Apr;39(1):43–56. doi: 10.1016/s0022-5320(72)80005-4. [DOI] [PubMed] [Google Scholar]
- Martin G. M., Sprague C. A., Epstein C. J. Replicative life-span of cultivated human cells. Effects of donor's age, tissue, and genotype. Lab Invest. 1970 Jul;23(1):86–92. [PubMed] [Google Scholar]
- Packer L., Deamer D. W., Heath R. L. Regulation and deterioration of structure in membranes. Adv Gerontol Res. 1967;2:77–120. [PubMed] [Google Scholar]
- Packer L., Smith J. R. Extension of the lifespan of cultured normal human diploid cells by vitamin E. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4763–4767. doi: 10.1073/pnas.71.12.4763. [DOI] [PMC free article] [PubMed] [Google Scholar]
- RUECKERT R. R., MUELLER G. C. Effect of oxygen tension on HeLa cell growth. Cancer Res. 1960 Jul;20:944–949. [PubMed] [Google Scholar]
- Rheinwald J. G., Green H. Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell. 1975 Nov;6(3):331–343. doi: 10.1016/s0092-8674(75)80001-8. [DOI] [PubMed] [Google Scholar]
- Robbins E., Levine E. M., Eagle H. Morphologic changes accompanying senescence of cultured human diploid cells. J Exp Med. 1970 Jun 1;131(6):1211–1222. doi: 10.1084/jem.131.6.1211. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ryan J. M., Sharf B. B., Cristofalo V. J. The influence of culture medium volume on cell density and lifespan of human diploid fibroblasts. Exp Cell Res. 1975 Mar 15;91(2):389–392. doi: 10.1016/0014-4827(75)90119-6. [DOI] [PubMed] [Google Scholar]
- TAPPEL A. L. FREE-RADICAL LIPID PEROXIDATION DAMAGE AND ITS INHIBITION BY VITAMIN E AND SELENIUM. Fed Proc. 1965 Jan-Feb;24:73–78. [PubMed] [Google Scholar]
- WARBURG O. On the origin of cancer cells. Science. 1956 Feb 24;123(3191):309–314. doi: 10.1126/science.123.3191.309. [DOI] [PubMed] [Google Scholar]
- Williamson A. R., Askonas B. A. Senescence of an antibody-forming cell clone. Nature. 1972 Aug 11;238(5363):337–339. doi: 10.1038/238337a0. [DOI] [PubMed] [Google Scholar]