Jaroch Karol, Taczyńska Paulina, Czechowska Marta, Bogusiewicz Joanna, Łuczykowski Kamil, Burlikowska Katarzyna, Bojko Barbara
Department of Pharmacodynamics and Molecular Pharmacology, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toruń Poland, 85-089, Bydgoszcz, Poland.
J Pharm Anal. 2021 Oct;11(5):667-674. doi: 10.1016/j.jpha.2021.03.005. Epub 2021 Mar 24.
Solid phase microextraction (SPME) in combination with high-resolution mass spectrometry was employed for the determination of metabolomic profile of mouse melanoma growth within in vitro 2D, in vitro 3D, and in vivo models. Such multi-model approach had never been investigated before. Due to the low-invasiveness of SPME, it was possible to perform time-course analysis, which allowed building time profile of biochemical reactions in the studied material. Such approach does not require the multiplication of samples as subsequent analyses are performed from the very same cell culture or from the same individual. SPME already reduces the number of animals required for experiment; therefore, it is with good concordance with the 3Rs rule (replacement, reduction, and refinement). Among tested models, the largest number of compounds was found within the in vitro 2D cell culture model, while in vivo and in vitro 3D models had the lowest number of detected compounds. These results may be connected with a higher metabolic rate, as well as lower integrity of the in vitro 2D model compared to the in vitro 3D model resulting in a lower number of compounds released into medium in the latter model. In terms of in vitro-in vivo extrapolation, the in vitro 2D model performed more similar to in vivo model compared to in vitro 3D model; however, it might have been due to the fact that only compounds secreted to medium were investigated. Thus, in further experiments to obtain full metabolome information, the intraspheroidal assessment or spheroid dissociation would be necessary.
采用固相微萃取（SPME）结合高分辨率质谱法测定小鼠黑色素瘤在体外二维、体外三维和体内模型中的代谢组学特征。这种多模型方法此前从未被研究过。由于SPME的侵入性低，因此可以进行时间进程分析，从而构建所研究材料中生化反应的时间图谱。这种方法不需要增加样本数量，因为后续分析是在同一细胞培养物或同一个体上进行的。SPME已经减少了实验所需的动物数量；因此，它与3R原则（替代、减少和优化）高度一致。在测试的模型中，在体外二维细胞培养模型中发现的化合物数量最多，而在体内和体外三维模型中检测到的化合物数量最少。这些结果可能与较高的代谢率有关，也与体外二维模型相比体外三维模型的完整性较低有关，导致后者模型中释放到培养基中的化合物数量较少。就体外-体内外推而言，与体外三维模型相比，体外二维模型与体内模型的表现更为相似；然而，这可能是因为只研究了分泌到培养基中的化合物。因此，在进一步的实验中，为了获得完整的代谢组信息，有必要进行球体内评估或球体解离。
