Adipose-derived stem cells (ASCs) and dedifferentiated fat (DFAT) cells are alternative cell sources in tissue engineering and regeneration because they are easily obtained and exhibit multilineage differentiation. However, aging may attenuate their regenerative potential and metabolic functions. Reports characterizing DFAT cells derived from aging donors are rare, and comparisons of DNA methylation profiles between aging ASCs and DFAT cells are poorly understood. Therefore, this study aimed to characterize DFAT cells relative to ASCs derived from aging subjects and compare the DNA methylation profiles of four adipogenic genes in these cells. ASCs and DFAT cells from aging donors exhibited characteristics similar to those of stem cells, including colony formation, proliferation, and multilineage differentiation abilities. However, compared with ASCs, DFAT cells exhibited increased proliferation, smooth muscle actin alpha (SMA-α) expression and decreased cellular senescence. DNA methylation profiling of ASCs and DFAT cells by combined bisulfite restriction analysis (COBRA) demonstrated hypermethylation patterns in three potent adipogenic genes—peroxisome proliferator-activated receptor gamma 2 (PPARγ2), fatty acid-binding protein 4 (FABP4), and lipoprotein lipase (LPL)—but hypomethylation of CCAAT/enhancer binding protein alpha (C/EBPα) in the aging group. Statistically significant differences were observed between the aging group and the young group. Epigenetic regulation maintains the stability of ASCs and DFAT cells in an age-dependent manner. Our findings suggested that although the DNA methylation patterns of three adipogenic genes correlated with hypermethylation and aging, ASCs and DFAT cells exhibited cellular stability and several stem cell characteristics, offering further opportunities for personalized regeneration and energy maintenance by adipogenesis during aging.

脂肪干细胞 (ASC) 和去分化脂肪 (DFAT) 细胞是组织工程和再生中的替代细胞来源，因为它们很容易获得并表现出多向分化。然而，衰老可能会削弱它们的再生潜力和代谢功能。表征来自衰老供体的 DFAT 细胞的报告很少见，并且对衰老 ASC 和 DFAT 细胞之间的 DNA 甲基化谱的比较知之甚少。因此，本研究旨在描述 DFAT 细胞相对于来自衰老受试者的 ASC 的特征，并比较这些细胞中四种脂肪形成基因的 DNA 甲基化谱。来自衰老供体的 ASC 和 DFAT 细胞表现出与干细胞相似的特征，包括集落形成、增殖和多向分化能力。然而，与 ASC 相比，DFAT 细胞表现出增殖增加、平滑肌肌动蛋白 α (SMA-α) 表达和细胞衰老减少。通过联合亚硫酸氢盐限制性分析 (COBRA) 对 ASC 和 DFAT 细胞的 DNA 甲基化谱分析显示了三种有效的脂肪生成基因——过氧化物酶体增殖物激活受体 γ2 (PPARγ2)、脂肪酸结合蛋白 4 (FABP4) 和脂蛋白脂肪酶中的高甲基化模式。 LPL)——但在衰老组中 CCAAT/增强子结合蛋白 α (C/EBPα) 的低甲基化。在老年组和年轻组之间观察到统计学上的显着差异。表观遗传调控以年龄依赖性方式维持 ASC 和 DFAT 细胞的稳定性。