Oxford Centre for Diabetes, Endocrinology and Metabolism, Radcliffe Department of Medicine, University of Oxford, Churchill Hospital, Headington, UK.
NIHR Oxford Biomedical Research Centre, OUH Foundation Trust, Oxford, UK.
Adipocyte. 2023 Dec;12(1):2179339. doi: 10.1080/21623945.2023.2179339.
While there is no standardized protocol for the differentiation of human adipocytes in culture, common themes exist in the use of supra-physiological glucose and hormone concentrations, and an absence of exogenous fatty acids. These factors can have detrimental effects on some aspects of adipogenesis and adipocyte function. Here, we present methods for modifying the adipogenic differentiation protocol to overcome impaired glucose uptake and insulin signalling in human adipose-derived stem cell lines derived from the stromal vascular fraction of abdominal and gluteal subcutaneous adipose tissue. By reducing the length of exposure to adipogenic hormones, in combination with a physiological glucose concentration (5 mM), and the provision of exogenous fatty acids (reflecting typical dietary fatty acids), we were able to restore early insulin signalling events and glucose uptake, which were impaired by extended use of hormones and a high glucose concentration, respectively. Furthermore, the addition of exogenous fatty acids greatly increased the storage of triglycerides and removed the artificial demand to synthesize all fatty acids by lipogenesis. Thus, modifying the adipogenic cocktail can enhance functional aspects of human adipocytes and is an important variable to consider prior to investigations into adipocyte biology.
虽然目前还没有标准化的方案来在培养条件下区分人类脂肪细胞，但是在使用超生理浓度的葡萄糖和激素，以及不添加外源性脂肪酸方面，存在一些共识。这些因素可能对脂肪生成和脂肪细胞功能的某些方面产生不利影响。在这里，我们介绍了一些修改脂肪生成分化方案的方法，以克服来源于腹部和臀部皮下脂肪组织基质血管部分的人脂肪源性干细胞系中葡萄糖摄取和胰岛素信号受损的问题。通过缩短暴露于脂肪生成激素的时间，结合生理浓度的葡萄糖（5mM），并提供外源性脂肪酸（反映典型的膳食脂肪酸），我们能够恢复早期胰岛素信号事件和葡萄糖摄取，而这两者分别受到激素和高葡萄糖浓度的延长作用而受损。此外，添加外源性脂肪酸大大增加了甘油三酯的储存，并通过脂肪生成去除了合成所有脂肪酸的人工需求。因此，修改脂肪生成混合物可以增强人类脂肪细胞的功能方面，并且是在研究脂肪细胞生物学之前需要考虑的一个重要变量。
