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New Approaches to Characterization of Aging Human Fibroblasts at Individual Cell Level

  • Youji Mitsui
  • Koji Matsuoka
  • Shinichi Aizawa
  • Koichi Noda
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 129)

Summary

Considering the heterogeneity of cell populations and other critcal problems in mass cultured senescent human fibroblasts, we proposed several new approaches for studying true cellular aging, as follows.
  1. 1)

    To establish a correlation among aging indexes at the individual cell level, we demonstrated a relationship among 3H-thymidine incorporation activity, nuclear size, cell volume, and DNA contents at individual cell level.

     
  2. 2)

    To fractionate homogenous cell populations and examine the relationship between their life spans and aging indexes, we separated human cells into relatively homogenous populations by the sedimentation velocity method, and found that life span of fractionated cells was almost identical among various fractions, irrespective of their great differences in cell volume and 3H-thymidine incorporation indexes. This suggests that some aging indexes, such as cell volumes and 3H-thymidine incorporation activity, are reversible cell properties, and are not specific properties at the individual cell level.

     
  3. 3)

    To find age-specific cell properties in fractionated senescent cell populations, we performed a quantitative analysis with an image analyzer on electromicroscopic pictures of fractionated small and large cell populations from young and senescent cultures. We discovered that the differences between young and senescent cultures in nuclear size, mitochonria size and number of lysosomes are due solely to the presence of large cells in senescent cultures, and that an increase in the constricted endoplasmic reticulum is a common phenotype of fractionated senescent cell populations. This suggests that there is some loss of function in endoplasmic reticulum or changes in the membrane system.

     
  4. 4)

    To examine age-specific changes in membrane system at the individual cell level, we found that the amount of absorption of concanavalin A-bound red blood cells to fibroblast surfaces increases linearly with in vitro passage. Further examination of individual cells indicated that this passage-related change in surface membrane is not dependent on cell cycle phase, surface area, or metabolic age, but certainly is dependent upon division age.

     

Finally, these findings should be confirmed by using cells from human donors of different ages.

We conclude that our approaches are important for elucidating the true mechanisms of cellular aging.

Keywords

Senescent Cell Nuclear Size Human Diploid Fibroblast Unlabeled Cell Aging Index 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    L. Hayflick, The limited number of in vitro lifetime of human diploid cell strains, Exp. Cell Res. 37:614, 1965.PubMedCrossRefGoogle Scholar
  2. 2.
    G. M. Martin, C. A. Sprague, and C. J. Epstein, Replicative lifespan of cultivated human cells: Effect of donor age, tissue, and genotype, Lab. Invest. 23:86, 1970.PubMedGoogle Scholar
  3. 3.
    E. L. Schneider and Y. Mitsui, The relationship between in vitro cellular aging and in vivo human age, Proc. Natl. Acad. Sci. USA 73:3584, 1976.PubMedCrossRefGoogle Scholar
  4. 4.
    V. J. Cristofalo, Thymidine labelling index as a criterion of aging in vitro, Gerontology 22:9, 1976.PubMedCrossRefGoogle Scholar
  5. 5.
    A. Macieira-Coelho, Kinetics of the proliferation of human fibroblasts during their lifespan in vitro, Mech. Ageing Dev. 5:45, 1977.Google Scholar
  6. 6.
    Y. Mitsui and E. L. Schneider, Relationship between cell replication and volume in senescent human diploid fibroblasts, Mech. Ageing Dev. 5:45, 1976.PubMedCrossRefGoogle Scholar
  7. 7.
    Y. Mitsui and E. L. Schneider, Increased nuclear sizes in senescent human diploid fibroblast cultures, Exp. Cell Res. 100:147, 1976.PubMedCrossRefGoogle Scholar
  8. 8.
    Y. Mitsui and E. L. Schneider, Characterization of fractionated human diploid fibroblast cell population, Exp. Cell Res. 103:23, 1976.PubMedCrossRefGoogle Scholar
  9. 9.
    J. J. Wolosewick and R. K. Porter, Observations on the morphological heterogeneity of WI-38, Am. J. Anat. 149:197, 1977.PubMedCrossRefGoogle Scholar
  10. 10.
    L. N. Kapp and R. R. Klevecz, The cell cycle of low passage and high passage human diploid fibroblasts, Exp. Cell Res. 101:154, 1976.PubMedCrossRefGoogle Scholar
  11. 11.
    Y. Mitsui, S. Aizawa, and K. Matsuoka, The relation of cell nuclei and surface membranes to the capacity of cell proliferation in human diploid fibroblasts, in: “Proceedings of XIth International Congress of Gerontology,” Excepta Medica, The Netherlands, p. 149, 1979.Google Scholar
  12. 12.
    S. Aizawa, Y. Mitsui, and F. Kurimoto, Cell surface changes accompanying aging in human diploid fibroblasts. II. Two types of age-related changes revealed by Concanavalin A-mediated red blood cell adsorption, Exp. Cell Res., in press.Google Scholar
  13. 13.
    H. Sakagomi, Y. Mitsui, S. Murota, and Y. Yamada, Two dimensional electrophoretic analysis of nuclear acidic proteins in senescent human fibroblasts, Cell Struc. Funct. 4:215, 1979.CrossRefGoogle Scholar
  14. 14.
    Y. Mitsui, H. Sakagami, S. Murota, and Y. Yamada, Age-related decline in Hi histone fraction in human diploid fibroblast cultures, Exp. Cell Res., in press.Google Scholar

Copyright information

© Plenum Press, New York 1980

Authors and Affiliations

  • Youji Mitsui
    • 1
  • Koji Matsuoka
    • 1
  • Shinichi Aizawa
    • 2
  • Koichi Noda
    • 3
  1. 1.Laboratory of PharmacologyTokyo Metropolitan Institute of GerontologyItabashiku, Tokyo-173Japan
  2. 2.Laboratory of NutritionTokyo Metropolitan Institute of GerontologyItabashiku, Tokyo-173Japan
  3. 3.Laboratory of Ultrastructure ResearchTokyo Metropolitan Institute of GerontologyItabashiku, Tokyo-173Japan

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