Hypertrophied human adipocyte spheroids as in vitro model of weight gain and adipose tissue dysfunction
Hypertrophied human adipocyte spheroids as in vitro model of weight gain and adipose tissue dysfunction
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DOI：
10.1113/jp281445
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发表时间：
2021-09-06
影响因子：
5.5
通讯作者：
Hagberg, Carolina E.
中科院分区：
文献类型：
--
作者：
Ioannidou, Anna;Alatar, Shemim;Hagberg, Carolina E.
脂肪细胞肥大是肥胖的一个重要特征，与胰岛素抵抗和代谢性疾病有关。由于缺乏具有代表性形态的人类细胞模型，脂肪细胞肥大的原因和后果在体外研究仍然很困难。（人单房血管化脂肪细胞球状体），对于具有更单室形态的人脂肪细胞的分化和培养，是获得具有大脂滴的体外分化脂肪细胞的关键HUVAS球状体的脂质处理可以进一步脂肪细胞扩大并诱导细胞功能障碍，模拟体重增加的体内效应。该模型将允许更广泛的研究团体对影响人类脂肪细胞分化和生长的因素进行机制研究，增加了我们对肥胖如何发展以及为什么它对全身代谢具有如此有害的后果的理解。肥胖流行率的上升迫切需要新的和改进的方法来研究人类脂肪细胞和体重增加在肥胖中的致病作用。体外尽管将脂肪祖细胞培养为3D结构的优势已得到证实，但大多数研究继续使用传统的2D培养物，这导致小的多室脂肪细胞具有较差的代表性。我们假设为分化中的前脂肪细胞提供血管生长龛将模拟体内脂肪形成并促进其分化为单室脂肪细胞。在这里，我们提出了HUVAS（人单室血管化脂肪细胞球体），一个简单的，易于应用的培养协议，允许分化的人脂肪细胞与更单室形态和更大的脂滴比以前的协议。此外，我们提供了一个协议，在体外诱导脂肪细胞扩大，导致更大的脂滴和脂肪细胞功能障碍的几个关键特征的发展，包括改变脂肪因子分泌，受损的脂解和胰岛素抵抗。总之，我们的HUVAS模型提供了一个改进的培养系统，用于研究体重增加期间引起人体脂肪组织代谢功能障碍和炎症的细胞和分子机制。
Key points Adipocyte enlargement is a key feature of obesity and associated with insulin resistance and metabolic disease The cause and consequences of adipocyte enlargement have remained hard to study in vitro due to a lack of human cell models with representative morphology This paper provides an easily set up spheroid culture method, HUVAS (human unilocular vascularized adipocyte spheroids), for the differentiation and culturing of human adipocytes with a more unilocular morphology We show that providing adipocyte progenitors with a vascular differentiation niche is key for achieving in vitro differentiated adipocytes with large lipid droplets Lipid treatment of the HUVAS spheroids can further adipocyte enlargement and induce cellular dysfunction, mimicking the in vivo effects of weight gain The model will allow a wider research community to perform mechanistic studies of the factors impacting on human adipocyte differentiation and growth, increasing our understanding of how obesity develops and why it has such detrimental consequences on whole body metabolism The rise in obesity prevalence has created an urgent need for new and improved methods to study human adipocytes and the pathogenic effects of weight gain in vitro. Despite the proven advantage of culturing adipocyte progenitors as 3D structures, the majority of studies continue to use traditional 2D cultures which result in small, multilocular adipocytes with poor representability. We hypothesized that providing differentiating pre-adipocytes with a vascular growth niche would mimic in vivo adipogenesis and improve the differentiation into unilocular adipocytes. Here we present HUVAS (human unilocular vascularized adipocyte spheroids), a simple, easily applicable culture protocol that allows for the differentiation of human adipocytes with a more unilocular morphology and larger lipid droplets than previous protocols. Moreover, we offer a protocol for inducing adipocyte enlargement in vitro, resulting in larger lipid droplets and development of several key features of adipocyte dysfunction, including altered adipokine secretion, impaired lipolysis and insulin resistance. Taken together, our HUVAS model offers an improved culture system for studying the cellular and molecular mechanisms causing metabolic dysfunction and inflammation in human adipose tissue during weight gain.