The State Key Lab of Analytical Chemistry for Life Science, Chemistry and Biomedicine Innovation Center (ChemBIC), School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210093, P. R. China.
Anal Chem. 2023 Jul 11;95(27):10221-10230. doi: 10.1021/acs.analchem.3c00479. Epub 2023 Jun 26.
Spatial metabolomic analysis of individual tumor spheroids can help investigate metabolic rearrangements in different cellular regions of a spheroid. In this work, a nanocapillary-based electrospray ionization mass spectroscopy (ESI-MS) method is established that could realize the spatial sampling of cellular components in different regions of a single living tumor spheroid and the subsequent MS analysis for a metabolic study. During the penetration of the nanocapillary into the spheroid for sampling, this "wound surface" at the outer layer of the spheroid takes only 0.1% of the whole area that maximally maintains the cellular activity inside the spheroid for the metabolic analysis. Using the ESI-MS analysis, different metabolic activities in the inner and outer (upper and lower) layers of a single spheroid are revealed, giving a full investigation of the metabolic heterogeneity inside one living tumor spheroid for the first time. In addition, the metabolic activities between the outer layer of the spheroid and two-dimensional (2D)-cultured cells show obvious differences, which suggests more frequent cell-cell and cell-extracellular environment interactions during the culture of the spheroid. This observation not only establishes a powerful tool for the in situ spatial analysis of the metabolic heterogeneity in single living tumor spheroids but also provides molecular information to elucidate the metabolic heterogeneity in this three-dimensional (3D)-cultured cell model.
单细胞球的空间代谢组学分析有助于研究细胞球不同区域的代谢重排。在这项工作中，建立了一种基于纳米毛细管的电喷雾电离质谱（ESI-MS）方法，该方法能够实现对单个活肿瘤细胞球不同区域的细胞成分进行空间采样，并对代谢研究进行后续的 MS 分析。在纳米毛细管进入细胞球进行采样的过程中，细胞球外层的“创面”仅占细胞球总面积的 0.1%，最大程度地保持了细胞球内的细胞活性，用于代谢分析。通过 ESI-MS 分析，揭示了单个细胞球的内、外层（上、下）之间不同的代谢活性，首次对一个活肿瘤细胞球内的代谢异质性进行了全面研究。此外，细胞球外层与二维（2D）培养细胞之间的代谢活性存在明显差异，这表明在细胞球培养过程中细胞-细胞和细胞-细胞外环境之间的相互作用更为频繁。这一观察结果不仅为单活肿瘤细胞球内代谢异质性的原位空间分析建立了一种强大的工具，而且为阐明该三维（3D）培养细胞模型中的代谢异质性提供了分子信息。
