CYTOLOGICAL AND CYTOGENETIC INVESTIGATIONS ON CELLULAR AGING IN HUMANS
Abstract
In vitro aging of human fibroblasts (W138) was systematically investigated by monitoring cumulative population doublings (CPD), average number of nucleoli, and nuclear and nucleolar volumes of cells grown in five media (Eagle's Basal Medium or BME, Medium 199 or M199, McCoy's 5A, Minimal Essential Medium or MEM, and HEPES-buffered Minimal Essential Medium or HMEM). It was found that the media are not interchangeable since cells grown in the five media were not identical in terms of all the parameters studied. Therefore, it is recommended that one standard culture system be utilized in using the in vitro system as a model for cellular aging of human diploid fibroblasts. Either BME, MEM or HMEM are suitable choices. Overall, during the in vitro aging process in all the media there was a linear increase in nuclear volume and average and total nucleolar volume of cells. We found a weak positive relationship between abnormal (irregular) nuclear morphology and cellular aging and a strong negative linear relationship between the average number of nucleoli and cellular senescence. M199 was very dissimilar to the other media studied. The uniqueness of M199 appeared to be due, in part, to the inclusion of Tween 80, a solubilizing detergent, in its formulation. This study also examined the effects of paramomycin (PM), an antibiotic, on in vitro cellular aging. It was found that paramomycin induces premature aging in that PM treated cells have a reduced lifespan, but are similar to control cells in terms of number of nucleoli, and nuclear and nucleolar volumes. This study also utilized lymphocyte cultures from humans of varying ages. Analysis of the frequency of cells with 44-47 chromosomes showed no differences between older (over 50 years of age) and younger subjects. Similarly there was no difference between the older and younger populations in terms of D-G satellite association. The Centromeric Separation Index (CSI) of older and younger subjects was found to differ significantly for chromosomes 6, X, 11, and 19. In the elderly, chromosomes 6 and X show an increased CSI while chromosomes 11 and 19 show a decreased CSI.
Subject Area
Genetics
Recommended Citation
WEINSTEIN, MARTHA E, "CYTOLOGICAL AND CYTOGENETIC INVESTIGATIONS ON CELLULAR AGING IN HUMANS" (1985). ETD Collection for Fordham University. AAI8521401.
https://research.library.fordham.edu/dissertations/AAI8521401
