Capturing Small Molecule Communication Between Tissues and Cells Using Imaging Mass Spectrometry
Capturing Small Molecule Communication Between Tissues and Cells Using Imaging Mass Spectrometry
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DOI：
10.3791/59490
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发表时间：
2019-04-01
影响因子：
1.2
通讯作者：
Sanchez, Laura M.
中科院分区：
文献类型：
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作者：
Zink, Katherine E.;Dean, Matthew;Sanchez, Laura M.
成像质谱（IMS）已常规应用于三种类型的样品：组织切片，球体和微生物菌落。这些样品类型已经使用基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）进行分析，以可视化蛋白质，脂质和代谢物在感兴趣的生物样品中的分布。我们开发了一种新的样品制备方法，结合了之前三种应用的优势，通过将哺乳动物细胞培养物播种到琼脂糖中，与健康组织共培养，然后将样品干燥，从而解决了一种尚未开发的方法，用于识别癌症中的化学通讯。哺乳动物的组织和细胞是近距离共培养的，通过组织和细胞之间的扩散使化学物质得以交流。在特定的时间点，琼脂糖为基础的样品干燥的方式与为IMS分析准备的微生物菌落相同。我们的方法旨在模拟源自输卵管的高级别浆液性卵巢癌在转移过程中与卵巢的相互作用。优化样品制备，鉴定去甲肾上腺素是卵巢微环境中的关键化学成分。这种新开发的方法可以应用于其他需要了解相邻细胞或组织之间化学通讯的生物系统。
Imaging mass spectrometry (IMS) has routinely been applied to three types of samples: tissue sections, spheroids, and microbial colonies. These sample types have been analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to visualize the distribution of proteins, lipids, and metabolites across the biological sample of interest. We have developed a novel sample preparation method that combines the strengths of the three previous applications to address an underexplored approach for identifying chemical communication in cancer, by seeding mammalian cell cultures into agarose in coculture with healthy tissues followed by desiccation of the sample. Mammalian tissue and cells are cocultured in close proximity allowing chemical communication via diffusion between the tissue and cells. At specific time points, the agarose-based sample is dried in the same manner as microbial colonies prepared for IMS analysis. Our method was developed to model the communication between high grade serous ovarian cancer derived from the fallopian tube as it interacts with the ovary during metastasis. Optimization of the sample preparation resulted in the identification of norepinephrine as a key chemical component in the ovarian microenvironment. This newly developed method can be applied to other biological systems that require an understanding of chemical communication between adjacent cells or tissues.