Olmedo-Suárez Miguel Angel, Sánchez-Ramírez Edgar, Del Toro-Rios Ximena, Noriega Lilia G, Aguilar-Arnal Lorena
Departamento de Biología Celular y Fisiología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Mexico City, Mexico.
Departamento de Fisiología de la Nutrición, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, Mexico.
Methods Mol Biol. 2025;2938:89-102. doi: 10.1007/978-1-0716-4607-6_10.
Human mesenchymal stem/stromal cells (hMSCs) constitute a primary cell source in adipogenesis, playing a pivotal role in the generation of new adipocytes. This process is accompanied by substantial metabolic shifts in hMSCs, notably characterized by a transition from glycolytic to oxidative metabolism. Monitoring metabolic alterations during adipogenic differentiation can be achieved through the assessment of extracellular flux parameters, such as the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). In this study, we provide a comprehensive protocol for differentiating hMSCs into the adipogenic lineage, with an emphasis on tracking differentiation stages through extracellular flux measurements. We determined the cellular metabolic status throughout the adipogenic differentiation by assessing OCR and ECAR to gauge mitochondrial respiration and glycolytic activity, respectively, using a Seahorse XFe96 extracellular flux analyzer.
人间充质干/基质细胞（hMSCs）是脂肪生成的主要细胞来源，在新脂肪细胞的生成中起关键作用。这一过程伴随着hMSCs显著的代谢转变，其显著特征是从糖酵解代谢转变为氧化代谢。通过评估细胞外通量参数，如耗氧率（OCR）和细胞外酸化率（ECAR），可以监测脂肪生成分化过程中的代谢变化。在本研究中，我们提供了一个将hMSCs分化为脂肪生成谱系的综合方案，重点是通过细胞外通量测量来跟踪分化阶段。我们使用Seahorse XFe96细胞外通量分析仪，通过评估OCR和ECAR分别测量线粒体呼吸和糖酵解活性，从而确定脂肪生成分化过程中的细胞代谢状态。
