Texas Pregnancy and Life-Course Health Research Center, Department of Animal Science, University of Wyoming, Laramie, WY, USA.
Barshop Institute for Longevity and Aging Studies and Department of Molecular Medicine, The University of Texas Health Science Center At San Antonio, 7703 Floyd Curl Dr, San Antonio, TX, 78229, USA.
Geroscience. 2021 Aug;43(4):2067-2085. doi: 10.1007/s11357-021-00395-1. Epub 2021 Jun 5.
Aging is associated with progressive loss of cellular homeostasis resulting from intrinsic and extrinsic challenges. Lack of a carefully designed, well-characterized, precise, translational experimental model is a major limitation to understanding the cellular perturbations that characterize aging. Here, we tested the feasibility of primary fibroblasts isolated from nonhuman primates (baboons) as a model of cellular resilience in response to homeostatic challenge. Using a real-time live-cell imaging system, we precisely defined a protocol for testing effects of prooxidant compounds (e.g., hydrogen peroxide (HO), paraquat), thapsigargin, dexamethasone, and a low glucose environment on cell proliferation in fibroblasts derived from baboons across the life course (n = 11/sex). Linear regression analysis indicated that donor age significantly reduced the ability of cells to proliferate following exposure to HO (50 and 100 µM) and paraquat (100 and 200 µM) challenges in cells from males (6.4-21.3 years; average lifespan 21 years) but not cells from females (4.3-15.9 years). Inhibitory effects of thapsigargin on cell proliferation were dependent on challenge duration (2 vs 24 h) and concentration (0.1 and 1 µM). Cells from older females (14.4-15.9 years) exhibited greater resilience to thapsigargin (1 µM; 24 h) and dexamethasone (500 µM) challenges than did those from younger females (4.3-6.7 years). The cell proliferation response to low glucose (1 mM) was reduced with age in both sexes. These data indicate that donor's chronological age and sex are important variables in determining fibroblast responses to metabolite and other challenges.
衰老是由于内在和外在挑战导致细胞内稳态逐渐丧失的结果。缺乏精心设计、特征明确、精确、转化实验模型是理解老化特征的细胞扰动的主要限制。在这里，我们测试了从非人灵长类动物（狒狒）分离的原代成纤维细胞作为细胞响应内稳态挑战的弹性模型的可行性。使用实时活细胞成像系统，我们精确地定义了一种用于测试促氧化剂化合物（例如，过氧化氢（HO）、百草枯）、他普西龙、地塞米松和低糖环境对来自狒狒的成纤维细胞增殖影响的方案，该模型涵盖了整个生命过程（n=11/性别）。线性回归分析表明，供体年龄显著降低了细胞暴露于 HO（50 和 100µM）和百草枯（100 和 200µM）挑战后增殖的能力雄性细胞（6.4-21.3 岁；平均寿命 21 岁），但不影响雌性细胞（4.3-15.9 岁）。他普西龙对细胞增殖的抑制作用取决于挑战持续时间（2 小时与 24 小时）和浓度（0.1 和 1µM）。来自较年长雌性（14.4-15.9 岁）的细胞对他普西龙（1µM；24 小时）和地塞米松（500µM）挑战的弹性大于来自较年轻雌性（4.3-6.7 岁）的细胞。低糖（1mM）对细胞增殖的反应随年龄在两性中降低。这些数据表明，供体的年龄和性别是决定成纤维细胞对代谢物和其他挑战反应的重要变量。
