Effective stem cell therapy is dependent on the stem cell quality that is determined by their differentiation potential, impairment of which leads to poor engraftment and survival into the target cells. However, limitations in our understanding and the lack of reliable markers that can predict their maturation efficacies have hindered the development of stem cells as an effective therapeutic strategy. Our previous study identified CD10, a pro-adipogenic, depot-specific prospective cell surface marker of human adipose-derived stem cells (ASCs). Here, we aim to determine if CD10 can be used as a prospective marker to predict mature adipocyte quality and play a direct role in adipocyte maturation. We first generated 14 primary human subject-derived ASCs and stable immortalized CD10 knockdown and overexpression lines for 4 subjects by the lentiviral transduction system. To evaluate the role of CD10 in adipogenesis, the adipogenic potential of the human subject samples were scored against their respective CD10 transcript levels. Assessment of UCP1 expression levels was performed to correlate CD10 levels to the browning potential of mature ASCs. Quantitative polymerase chain reaction (qPCR) and Western blot analysis were performed to determine CD10-dependent regulation of various targets. Seahorse analysis of oxidative metabolism and lipolysis assay were studied. Lastly, as a proof-of-concept study, we used CD10 as a prospective marker for screening nuclear receptor ligands library. We identified intrinsic CD10 levels as a positive determinant of adipocyte maturation as well as browning potential of ASCs. Interestingly, CD10 regulates ASC’s adipogenic maturation non-canonically by modulating endogenous lipolysis without affecting the classical peroxisome proliferator-activated receptor gamma (PPARγ)-dependent adipogenic pathways. Furthermore, our CD10-mediated screening analysis identified dexamethasone and retinoic acid as stimulator and inhibitor of adipogenesis, respectively, indicating CD10 as a useful biomarker for pro-adipogenic drug screening. Overall, we establish CD10 as a functionally relevant ASC biomarker, which may be a prerequisite to identify high-quality cell populations for improving metabolic diseases.

中文翻译：

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CD10 标记非典型 PPARγ 独立脂肪细胞成熟和脂肪干细胞的褐变潜力


有效的干细胞治疗取决于干细胞的质量，而干细胞的质量由其分化潜力决定，分化潜力的损害会导致靶细胞的植入和存活不良。然而，我们理解的局限性以及缺乏可以预测其成熟功效的可靠标记物阻碍了干细胞作为有效治疗策略的发展。我们之前的研究发现了 CD10，这是一种人类脂肪干细胞 (ASC) 的促脂肪生成、储库特异性的前瞻性细胞表面标记物。在这里，我们的目的是确定CD10是否可以作为预测成熟脂肪细胞质量的前瞻性标志物并在脂肪细胞成熟中发挥直接作用。我们首先通过慢病毒转导系统为 4 名受试者生成了 14 个原代人类受试者来源的 ASC 和稳定的永生化 CD10 敲低和过表达系。为了评估 CD10 在脂肪形成中的作用，根据人类受试者样本各自的 CD10 转录物水平对其脂肪形成潜力进行评分。对 UCP1 表达水平进行评估，将 CD10 水平与成熟 ASC 的褐变潜力相关联。进行定量聚合酶链反应 (qPCR) 和蛋白质印迹分析以确定各种靶标的 CD10 依赖性调节。研究了海马氧化代谢分析和脂肪分解测定。最后，作为概念验证研究，我们使用 CD10 作为筛选核受体配体库的前瞻性标记。我们确定内在 CD10 水平是脂肪细胞成熟以及 ASC 褐变潜力的积极决定因素。 有趣的是，CD10 通过调节内源性脂肪分解来非规范地调节 ASC 的脂肪形成成熟，而不影响经典的过氧化物酶体增殖物激活受体 γ (PPARγ) 依赖性脂肪形成途径。此外，我们的 CD10 介导的筛选分析确定地塞米松和视黄酸分别作为脂肪生成的刺激剂和抑制剂，表明 CD10 作为促脂肪生成药物筛选的有用生物标志物。总体而言，我们将 CD10 确立为功能相关的 ASC 生物标志物，这可能是识别改善代谢疾病的高质量细胞群的先决条件。