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引用次数: 0
摘要
白色脂肪组织(WAT)是调节血糖和血脂水平、饱腹感和炎症的重要内分泌器官。在采用所述技术之前,原代白色脂肪细胞无法在体外稳定培养。缺乏可靠的原代培养模型阻碍了白脂肪代谢和药物开发方面的研究。我们开发了一种新型的白脂肪原代培养技术,称为 "夹层白脂肪组织"(SWAT)。SWAT 通过在脂肪基质细胞薄片之间夹入切碎的白脂肪组织,克服了脂肪细胞的自然浮力。由此产生的构建体在培养过程中至少可存活 8 周。SWAT 保持了完整的细胞外基质、细胞间接触和体内 WAT 条件下的物理压力;此外,SWAT 还保持了强大的转录谱、对外源化学信号的敏感性和整体组织功能。SWAT 是一种简单、可重复且有效的原代脂肪培养方法。它有可能成为一个广泛适用于脂肪细胞生理学、病理生理学、新陈代谢和药物开发研究的平台。
White adipose tissue (WAT) plays a crucial endocrine organ that regulates blood glucose and lipid levels, satiety, and inflammation. Before the described technique, primary white adipocytes could not be stably cultured in vitro. The lack of a reliable primary culture model impeded research in WAT metabolism and drug development. We have developed a novel technique for WAT primary culture called "sandwiched white adipose tissue" (SWAT). SWAT overcomes the natural buoyancy of adipocytes by sandwiching minced WAT between sheets of adipose-derived stromal cells. The resulting constructs are viable for at least 8 weeks in culture. SWAT maintains the intact extracellular matrix, cell-to-cell contacts, and physical pressures of in vivo WAT conditions; additionally, SWAT maintains a robust transcriptional profile, sensitivity to exogenous chemical signaling, and whole tissue function. SWAT represents a simple, reproducible, and effective method of primary adipose culture. Potentially, it is a broadly applicable platform for research in WAT physiology, pathophysiology, metabolism, and pharmaceutical development.
期刊介绍:
For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.