Herbers Elena, Patrikoski Mimmi, Wagner Anita, Jokinen Riikka, Hassinen Antti, Heinonen Sini, Miettinen Susanna, Peltoniemi Hilkka, Pirinen Eija, Pietiläinen Kirsi H
Obesity Research Unit, Research Program for Clinical and Molecular Metabolism, Faculty of Medicine, University of Helsinki, Helsinki, Finland.
Research Program for Clinical and Molecular Metabolism, Faculty of Medicine, University of Helsinki, Helsinki, Finland.
Stem Cells Int. 2022 Apr 5;2022:3308194. doi: 10.1155/2022/3308194. eCollection 2022.
Mitochondrial dysfunction in white adipose tissue is strongly associated with obesity and its metabolic complications, which are important health challenges worldwide. Human adipose-derived stromal/stem cells (hASCs) are a promising tool to investigate the underlying mechanisms of such mitochondrial dysfunction and to subsequently provide knowledge for the development of treatments for obesity-related pathologies. A substantial obstacle in using hASCs is that the key compounds for adipogenic differentiation increase mitochondrial uncoupling, biogenesis, and activity, which are the signature features of brown adipocytes, thus altering the white adipocyte phenotype towards brown-like cells. Additionally, commonly used protocols for hASC adipogenic differentiation exhibit high variation in their composition of media, and a systematic comparison of their effect on mitochondria is missing. Here, we compared the five widely used adipogenic differentiation protocols for their effect on metabolic and mitochondrial phenotypes to identify a protocol that enables differentiation of white adipocytes and can more faithfully recapitulate the white adipocyte phenotype observed in human adipose tissue. We developed a workflow that included functional assays and morphological analysis of mitochondria and lipid droplets. We observed that triiodothyronine- or indomethacin-containing media and commercially available adipogenic media induced browning during differentiation of white adipocytes. However, the differentiation protocol containing 1 M of the peroxisome proliferator-activated receptor gamma (PPAR) agonist rosiglitazone prevented the browning effect and would be proposed for adipogenic differentiation protocol for hASCs to induce a white adipocyte phenotype. Preserving the white adipocyte phenotype is a crucial step for the study of obesity and associated metabolic diseases, adipose tissue pathologies, such as lipodystrophies, possible therapeutic compounds, and basic adipose tissue physiology.
白色脂肪组织中的线粒体功能障碍与肥胖及其代谢并发症密切相关，而肥胖及其代谢并发症是全球范围内重要的健康挑战。人脂肪来源的基质/干细胞（hASC）是一种很有前景的工具，可用于研究此类线粒体功能障碍的潜在机制，并为开发肥胖相关病症的治疗方法提供知识。使用hASC的一个重大障碍是，成脂分化的关键化合物会增加线粒体解偶联、生物发生和活性，这些是棕色脂肪细胞的标志性特征，从而使白色脂肪细胞表型向类棕色细胞转变。此外，常用的hASC成脂分化方案在培养基组成上存在很大差异，并且缺少对它们对线粒体影响的系统比较。在这里，我们比较了五种广泛使用的成脂分化方案对代谢和线粒体表型的影响，以确定一种能够实现白色脂肪细胞分化并能更忠实地重现人脂肪组织中观察到的白色脂肪细胞表型的方案。我们开发了一种工作流程，包括对线粒体和脂滴的功能测定和形态分析。我们观察到，含三碘甲状腺原氨酸或吲哚美辛的培养基以及市售的成脂培养基在白色脂肪细胞分化过程中会诱导褐变。然而，含有1 M过氧化物酶体增殖物激活受体γ（PPAR）激动剂罗格列酮的分化方案可防止褐变效应，因此建议将其用于hASC的成脂分化方案以诱导白色脂肪细胞表型。保留白色脂肪细胞表型是研究肥胖及相关代谢疾病、脂肪组织病理学（如脂肪营养不良）、可能的治疗化合物以及基础脂肪组织生理学的关键步骤。
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