Simple flow cytometric method used to assess lipid accumulation in fat cells
Simple flow cytometric method used to assess lipid accumulation in fat cells
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DOI：
10.1194/jlr.d300028-jlr200
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发表时间：
2004-06-01
影响因子：
6.5
通讯作者：
Huang, YF
中科院分区：
文献类型：
--
作者：
Lee, YH;Chen, SY;Huang, YF
前脂肪细胞的成脂作用已被广泛用于研究药物对脂肪细胞转化的分子基础和作用。然而，在成脂诱导后，细胞对诱导剂的转化和脂肪积累的速度不同，这使得直接的分子和生化分析变得复杂。在这里，我们提出了一种简单而灵敏的方法，用来检测和量化细胞内的脂肪积聚。使用这种方法，我们检测到细胞质颗粒水平的升高与成脂转化后细胞内积累的脂肪水平的增加有很好的相关性。我们进一步证明了这种方法能够监测和量化复杂细胞群体中的脂肪细胞成熟，并识别和收集具有相似脂肪存储的脂肪细胞用于进一步分析。与现有的细胞质脂质染色和脂肪提取检测系统相比，流式细胞术具有明显的优势，可以作为一种强大的分析工具来监测化学物质和生物分子对脂肪细胞转化和成熟的影响。此外，结合细胞分选设备，我们的方法提供了一种简单而有效的方法来收集特定状态的脂肪细胞用于进一步分析。
Adipogenesis of preadipocytes in culture has been frequently used to study the molecular basis and effect of drugs on fat cell conversion. However, after adipogenic induction, cells respond to the inducing agent with various speeds of conversion and fat accumulation, which complicates direct molecular and biochemical analyses. Here we present a simple and sensitive method to detect and quantify fat accumulation inside cells by flow cytometry. Using this method, we detected elevated levels of cytoplasmic granularity that correlated well with an increased level of fat accumulated inside cells after adipogenic conversion. We further demonstrated the ability of this method to monitor and quantify fat cell maturation within a complex population of cells and to identify and collect the fat cells with similar fat storage for further analysis. Flow cytometry offers distinct advantages over existing detection systems for cytoplasmic lipid staining and lipid extraction and could represent a powerful analytical tool to monitor the effect of chemicals and biological molecules on fat cell conversion and maturation. Moreover, in combination with a cell sorting facility, our method offers a simple and efficient means of collecting fat cells of specific status for further analysis.