Maintenance of Chronological Aging Features in Culture of Normal Human Dermal Fibroblasts from Old Donors

Abstract

Chronological aging is defined as a time-dependent decline of tissue homeostasis which severely impacts skin. Understanding the mechanisms of skin aging is an active research area limited by the lack of relevant in vitro models. Being a component of aging, replicative or stress-induced senescence is repeatedly used to mimic skin aging in vitro, thus presenting only a partial view of the complexity of aging. Herein, we aimed to clarify whether primary normal human dermal fibroblasts retained age-related characteristics when cultured in 2D monolayer, and could be used as a relevant model for aging research. We compared three groups of fibroblasts isolated from different aged donors. We observed strongly decreased population doubling capacities, a reduced clonogenic ability, an impairment in extracellular matrix production together with modifications of respiratory metabolism with an increase in age. These disruptions were particularly marked when comparing fibroblasts isolated from old individuals (over 70 years old) to those isolated from young individuals (18–37 years old), while cells from middle-aged donors exhibited an intermediate profile. These alterations of cell features can be related to the signs of dermis aging, thus showing that cultured primary cells indeed retain some characteristics of the original tissue from which they were extracted.

Effects of Betulinic Acid on the Proliferation, Cellular Senescence, and Type 1 Interferon-Related Signaling Pathways in Human Dermal Fibroblasts

“…It is also used as an inducer of cellular senescence. 26 When we cultured NHDFs with etoposide for 48 h and then cultured them with a normal medium for an additional 1 week, we found an increase in (Figure 8C). In addition, BA treatment attenuated p21 expression, whereas etoposide-treated NHDFs exhibited a higher p21 expression than control cells (Figure 8D).…”

“…Etoposide is an anticancer drug that targets topoisomerase II. It is also used as an inducer of cellular senescence . When we cultured NHDFs with etoposide for 48 h and then cultured them with a normal medium for an additional 1 week, we found an increase in positive cells of SA-β-galactosidase activity (Figure A,B).…”

Protocol Optimization for Direct Reprogramming of Primary Human Fibroblast into Induced Striatal Neurons

“…Neurons are believed to have permanently blocked their capacity to proliferate once they are differentiated, being typically found in a quiescent state in the adult nervous system. Meanwhile, dermal fibroblasts obtained from young and middle-aged donors, an age when HD manifests, represent a population of actively dividing cells [ 38 ]. Moreover, the results from another study group have demonstrated that the attenuation of p53, in conjunction with cell cycle arrest at G1 obtained by serum starvation and an appropriate cell culture environment, increases the efficiency of the transdifferentiation of human fibroblasts to induced dopaminergic neurons [ 39 ].…”

Focus on the Contribution of Oxidative Stress in Skin Aging

“…Since it has been proposed that reticular fibroblasts are in a more advanced stage of differentiation than papillary fibroblasts, it seems plausible that the transition from the papillar to reticular state is related to the relentless terminal differentiation process of post-mitotic cells [ 202 , 203 ]. Another recent study proposed a specific feature of chronological age-related partial loss of dermal fibroblasts described as “loss of cellular identity” [ 204 ]. In detail, old papillary fibroblasts presented fewer papillary and more reticular gene expression profiles, while the reticular counterpart presented a less decided reticular gene expression profile [ 205 ].…”