Aim: Ceiling culture-derived preadipocytes (ccdPAs) and adipose-derived stem cells (ASCs) can be harvested from human subcutaneous fat tissue using the specific gravity method. Both cell types possess a similar spindle shape without lipid droplets. We previously reported that ccdPAs have a higher adipogenic potential than ASCs, even after a 7-week culture. We performed a genome-wide epigenetic analysis to examine the mechanisms contributing to the adipogenic potential differences between ccdPAs and ASCs. Materials and Methods: Methylation analysis of cytosines followed by guanine (CpG) using a 450K BeadChip was performed on human ccdPAs and ASCs isolated from three metabolically healthy females. Chromatin immunoprecipitation sequencing (ChIP-seq) was performed to evaluate trimethylation at lysine 4 of histone 3 (H3K4me3). Results: Unsupervised machine learning using t-distributed stochastic neighbor embedding (tSNE) to interpret 450,000-dimensional methylation assay data showed that the cells were divided into ASC and ccdPA groups. In KEGG pathway analysis of 1,543 genes with differential promoter CpG methylation, the peroxisome proliferator-activated receptor (PPAR) and adipocytokine signaling pathways ranked in the top 10 pathways. In the PPAR gamma gene, H3K4me3 peak levels were higher in ccdPAs than in ASCs, whereas promoter CpG methylation levels were significantly lower in ccdPAs than in ASCs. Similar differences in promoter CpG methylation were also seen in the fatty acid-binding protein 4 (FABP4) and leptin genes. Conclusion: We analyzed the epigenetic status of adipogenesis-related genes as a potential mechanism underlying the differences in adipogenic differentiation capability between ASCs and ccdPAs.

中文翻译：

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表观遗传修饰是源自人皮下脂肪组织的前脂肪细胞差异成脂潜力的基础

目的：可以使用比重法从人体皮下脂肪组织中收集天花板培养衍生的前脂肪细胞 (ccdPAs) 和脂肪衍生的干细胞 (ASCs)。两种细胞类型都具有相似的纺锤形，没有脂滴。我们之前曾报道 ccdPAs 比 ASCs 具有更高的成脂潜力，即使经过 7 周的培养。我们进行了全基因组表观遗传分析，以检查导致 ccdPA 和 ASC 之间成脂潜力差异的机制。材料和方法：使用 450K BeadChip 对从三名代谢健康女性中分离的人 ccdPA 和 ASC 进行胞嘧啶甲基化分析，然后是鸟嘌呤 (CpG)。进行染色质免疫沉淀测序 (ChIP-seq) 以评估组蛋白 3 (H3K4me3) 的赖氨酸 4 处的三甲基化。结果：使用 t 分布随机邻域嵌入 (tSNE) 来解释 450,000 维甲基化测定数据的无监督机器学习表明，细胞分为 ASC 和 ccdPA 组。在对具有差异启动子 CpG 甲基化的 1,543 个基因的 KEGG 通路分析中，过氧化物酶体增殖物激活受体 (PPAR) 和脂肪细胞因子信号通路排在前 10 名通路中。在 PPAR γ 基因中，ccdPAs 中的 H3K4me3 峰值水平高于 ASCs，而 ccdPAs 中的启动子 CpG 甲基化水平显着低于 ASCs。在脂肪酸结合蛋白 4 (FABP4) 和瘦素基因中也观察到启动子 CpG 甲基化的类似差异。结论：