Isolation and Culture of Human Mature Adipocytes Using Membrane Mature Adipocyte Aggregate Cultures (MAAC)
Isolation and Culture of Human Mature Adipocytes Using Membrane Mature Adipocyte Aggregate Cultures (MAAC)
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DOI：
10.3791/60485
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发表时间：
2020-02-01
影响因子：
1.2
通讯作者：
Boucher, Jeremie
中科院分区：
文献类型：
--
作者：
Alexandersson, Ida;Harms, Matthew J.;Boucher, Jeremie
白色脂肪组织（WAT）失调在胰岛素抵抗和2型糖尿病（T2 D）的发展中起着重要作用。为了开发T2D的新治疗方法，需要更生理相关的体外脂肪细胞模型。本研究描述了一种分离和培养成熟人脂肪细胞的新技术。该方法被称为MAAC（膜成熟脂肪细胞聚集体培养），与其他脂肪细胞体外模型相比，MAAC具有最接近新鲜分离的成熟脂肪细胞的成脂基因特征。使用MAAC，脂肪细胞可以从瘦和肥胖患者、不同的脂肪库培养，与不同的细胞类型共培养，并且重要的是，可以保持培养2周。还可以对MAAC进行功能实验，包括葡萄糖摄取、脂肪生成和脂解。此外，MAAC对不同的药理学激动作用有强烈的反应，可用于研究脂肪细胞表型的变化，包括白色脂肪细胞向棕色样脂肪细胞的转分化。
White adipose tissue (WAT) dysregulation plays a central role in development of insulin resistance and type 2 diabetes (T2D). To develop new treatments for T2D, more physiologically relevant in vitro adipocyte models are required. This study describes a new technique to isolate and culture mature human adipocytes. This method is entitled MAAC (membrane mature adipocyte aggregate culture), and compared to other adipocyte in vitro models, MAAC possesses an adipogenic gene signature that is the closest to freshly isolated mature adipocytes. Using MAAC, adipocytes can be cultured from lean and obese patients, different adipose depots, co-cultured with different cell types, and importantly, can be kept in culture for 2 weeks. Functional experiments can also be performed on MAAC including glucose uptake, lipogenesis, and lipolysis. Moreover, MAAC responds robustly to diverse pharmacological agonism and can be used to study adipocyte phenotypic changes, including the transdifferentiation of white adipocytes into brown-like fat cells.