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)


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.


Senescent Cell Nuclear Size Human Diploid Fibroblast Unlabeled Cell Aging Index 
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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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