Department of Pathology, University of Colorado Denver, Anschutz Medical Campus, Aurora, CO 80045, USA.
J Mammary Gland Biol Neoplasia. 2009 Dec;14(4):381-6. doi: 10.1007/s10911-009-9161-5. Epub 2009 Dec 2.
Analysis of genes and proteins involved in lipid biosynthesis in mammary epithelial cells (MECs) is complicated by the presence of adipose tissue in the mammary gland, which may be predominant in whole tissue lysates depending upon developmental stage. We have developed a method based on protocols used to establish primary mammary epithelial cell cultures that allows for analysis of MECs depleted of adipose. Adipose depletion yields enriched MECs that are suitable for gene expression profiling and protein analysis from a single mouse. Additionally, the phosphorylation of proteins is maintained, allowing investigation of signal transduction events. Application of this method to the analysis of MECs from genetically modified mice will aid in the identification of factors controlling tissue-specific events in the mammary gland. In contrast to other methods such as laser capture microdissection, the MEC enrichment method described here is performed using standard lab supplies, equipment, and techniques.
分析乳腺上皮细胞（MEC）中参与脂类生物合成的基因和蛋白质很复杂，因为乳腺中存在脂肪组织，根据发育阶段的不同，脂肪组织可能在整个组织裂解物中占主导地位。我们已经开发了一种基于建立原代乳腺上皮细胞培养物的方案的方法，该方法允许分析去除脂肪的 MEC。脂肪去除可产生富含 MEC 的细胞，适合从单个小鼠进行基因表达谱分析和蛋白质分析。此外，蛋白质的磷酸化得到维持，允许研究信号转导事件。将该方法应用于分析基因修饰小鼠的 MEC 将有助于鉴定控制乳腺组织特异性事件的因素。与其他方法（如激光捕获显微解剖）相比，这里描述的 MEC 富集方法使用标准实验室用品、设备和技术进行。
