Diet, Genomics and Immunology Laboratory, Beltsville Human Nutrition Research Center, Agricultural Research Service, United States Department of Agriculture, Beltsville, MD 20705, USA.
Division of Food Labeling & Standards, Office of Nutrition and Food Labeling, Center for Food Safety and Applied Nutrition, U.S. Food and Drug Administration, College Park, MD 20740, USA.
Int J Mol Sci. 2022 Oct 19;23(20):12534. doi: 10.3390/ijms232012534.
Adipose stem cells (ASCs) are reported to play a role in normal physiology as well as in inflammation and disease. The objective of this work was to elucidate inter-individual differences in growth, gene expression and response to inflammatory stimuli in ASCs from different donors. Human ASC1 (male donor) and ASC2 (female donor) were purchased from Lonza (Walkersville, MD). Cell proliferation was determined by the sulforhodamine B assay. After time-dependent treatment of ASCs with or without bacterial lipopolysaccharide (LPS), marker gene mRNAs for proliferation, steroid hormones, and xenobiotic and immune pathways were determined using RT-PCR, and secreted cytokine levels in media were measured using the Bio-Plex cytokine assay kit. ASCs from both donors expressed androgen receptors but not estrogen receptors. ASC2 had a 2-fold higher proliferation rate and a 6-fold higher level of proliferation marker Ki67 mRNA than ASC1. ASC2 exhibited significantly greater fold induction of TNF-α and CCL2 by LPS compared to ASC1. TNF-α and GM-CSF protein levels were also significantly higher in the LPS-induced ASC2 media, but IL-6 secretion was higher in the LPS-induced ASC1 media. Our findings suggest that inter-individual variability and/or possible sex differences exist in ASCs, which may serve as a key determinant to inflammatory responses of ASCs.
脂肪干细胞（ASCs）被报道在正常生理、炎症和疾病中发挥作用。本研究旨在阐明不同供体来源的 ASC 之间在生长、基因表达和对炎症刺激反应方面的个体间差异。人 ASC1（男性供体）和 ASC2（女性供体）购自 Lonza（马里兰州沃克斯维尔）。通过磺酰罗丹明 B 测定法测定细胞增殖。用或不用细菌脂多糖（LPS）对 ASC 进行时间依赖性处理后，使用 RT-PCR 测定增殖、甾体激素、外源性和免疫途径的标记基因 mRNA，使用 Bio-Plex 细胞因子检测试剂盒测定培养基中分泌的细胞因子水平。来自两个供体的 ASC 均表达雄激素受体，但不表达雌激素受体。与 ASC1 相比，ASC2 的增殖率高 2 倍，增殖标志物 Ki67 mRNA 水平高 6 倍。与 ASC1 相比，LPS 诱导的 ASC2 中 TNF-α 和 CCL2 的诱导倍数显著更高。LPS 诱导的 ASC2 培养基中的 TNF-α 和 GM-CSF 蛋白水平也明显更高，但 LPS 诱导的 ASC1 培养基中的 IL-6 分泌更高。我们的研究结果表明，ASCs 存在个体间变异性和/或可能的性别差异，这可能是 ASC 炎症反应的关键决定因素。
