Research Paper Volume 9, Issue 5 pp 1404—1413
Diffuse colonies of human skin fibroblasts in relation to cellular senescence and proliferation
- 1 Human Stem Cells Institute, Moscow, 119333, Russia
- 2 State Research Center - Burnasyan Federal Medical Biophysical Center of Federal Medical Biological Agency, Moscow, 123098, Russia
- 3 Blokhin Cancer Research Center, Moscow, 115478, Russia
- 4 Moscow Institute of Physics and Technology, Dolgoprudny, Moscow Region, 141700, Russia
- 5 Institute of Cell Biophysics, Russian Academy of Sciences, Pushchino, Moscow Region, 142290, Russia
- 6 Canadian Nuclear Laboratories, Chalk River, Ontario, K0J1J0, Canada
- 7 Semenov Institute of Chemical Physics, Russian Academy of Sciences, Moscow, 119991, Russia
received: January 29, 2017 ; accepted: May 11, 2017 ; published: May 16, 2017 ;https://doi.org/10.18632/aging.101240
How to Cite
Copyright: Zorin et al. This is an open‐access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Development of personalized skin treatment in medicine and skin care may benefit from simple and accurate evaluation of the fraction of senescent skin fibroblasts that lost their proliferative capacity. We examined whether enriched analysis of colonies formed by primary human skin fibroblasts, a simple and widely available cellular assay, could reveal correlations with the fraction of senescent cells in heterogenic cell population. We measured fractions of senescence associated β-galactosidase (SA-βgal) positive cells in either mass cultures or colonies of various morphological types (dense, mixed and diffuse) formed by skin fibroblasts from 10 human donors. Although the donors were chosen to be within the same age group (33-54 years), the colony forming efficiency of their fibroblasts (ECO-f) and the percentage of dense, mixed and diffuse colonies varied greatly among the donors. We showed, for the first time, that the SA-βgal positive fraction was the largest in diffuse colonies, confirming that they originated from cells with the least proliferative capacity. The percentage of diffuse colonies was also found to correlate with the SA-βgal positive cells in mass culture. Using Ki67 as a cell proliferation marker, we further demonstrated a strong inverse correlation (r=-0.85, p=0.02) between the percentage of diffuse colonies and the fraction of Ki67+ cells. Moreover, a significant inverse correlation (r=-0.94, p=0.0001) between the percentage of diffuse colonies and ECO-f was found. Our data indicate that quantification of a fraction of diffuse colonies may provide a simple and useful method to evaluate the extent of cellular senescence in human skin fibroblasts.