An optimised protocol for the investigation of insulin signalling in a human cell culture model of adipogenesis.

IF 3.5 4区 生物学 Q2 ENDOCRINOLOGY & METABOLISM Adipocyte Pub Date : 2023-12-01 DOI:10.1080/21623945.2023.2179339
Jonathan M Gamwell, Keanu Paphiti, Leanne Hodson, Fredrik Karpe, Katherine E Pinnick, Marijana Todorčević
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Abstract

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 de novo lipogenesis. Thus, modifying the adipogenic cocktail can enhance functional aspects of human adipocytes in vitro and is an important variable to consider prior to in vitro investigations into adipocyte biology.

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胰岛素信号在脂肪生成的人类细胞培养模型中研究的优化方案。
虽然没有在培养中分化人类脂肪细胞的标准化方案,但在使用超生理葡萄糖和激素浓度以及缺乏外源性脂肪酸方面存在共同的主题。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。在此,我们提出了修改脂肪分化方案的方法,以克服来源于腹部和臀大肌皮下脂肪组织基质血管部分的人类脂肪来源干细胞系中葡萄糖摄取和胰岛素信号传导受损的问题。通过减少暴露于脂肪生成激素的时间,结合生理葡萄糖浓度(5mM)和外源性脂肪酸(反映典型的膳食脂肪酸)的提供,我们能够恢复早期胰岛素信号事件和葡萄糖摄取,这两个事件分别因长时间使用激素和高葡萄糖浓度而受损。此外,添加外源性脂肪酸大大增加了甘油三酯的储存,并消除了通过从头脂肪生成合成所有脂肪酸的人为需求。因此,修饰成脂混合物可以在体外增强人类脂肪细胞的功能,并且是在体外研究脂肪细胞生物学之前需要考虑的一个重要变量。
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来源期刊
Adipocyte
Adipocyte Medicine-Histology
CiteScore
6.50
自引率
3.00%
发文量
46
审稿时长
32 weeks
期刊介绍: Adipocyte recognizes that the adipose tissue is the largest endocrine organ in the body, and explores the link between dysfunctional adipose tissue and the growing number of chronic diseases including diabetes, hypertension, cardiovascular disease and cancer. Historically, the primary function of the adipose tissue was limited to energy storage and thermoregulation. However, a plethora of research over the past 3 decades has recognized the dynamic role of the adipose tissue and its contribution to a variety of physiological processes including reproduction, angiogenesis, apoptosis, inflammation, blood pressure, coagulation, fibrinolysis, immunity and general metabolic homeostasis. The field of Adipose Tissue research has grown tremendously, and Adipocyte is the first international peer-reviewed journal of its kind providing a multi-disciplinary forum for research focusing exclusively on all aspects of adipose tissue physiology and pathophysiology. Adipocyte accepts high-profile submissions in basic, translational and clinical research.
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