School of Chemical, Materials, and Biomedical Engineering, University of Georgia, Athens, GA 30602, USA; Regenerative Bioscience Center, Rhodes Center for ADS, University of Georgia, Athens, GA 30602, USA.
School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA 30332, USA.
Cell Rep. 2024 Aug 27;43(8):114579. doi: 10.1016/j.celrep.2024.114579. Epub 2024 Aug 15.
Mesenchymal stem/stromal cell (MSC) therapies have had limited success so far in clinical trials due in part to heterogeneity in immune-responsive phenotypes. Therefore, techniques to characterize these properties of MSCs are needed during biomanufacturing. Imaging cell shape, or morphology, has been found to be associated with MSC immune responsivity-but a direct relationship between single-cell morphology and function has not been established. We used label-free differential phase contrast imaging and matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) to evaluate single-cell morphology and explore relationships with lipid metabolic immune response. In interferon gamma (IFN-γ)-stimulated MSCs, we found higher lipid abundances from the ceramide-1-phosphate (C1P), phosphatidylcholine (PC), LysoPC, and triglyceride (TAG) families that are involved in cell immune function. Furthermore, we identified differences in lipid signatures in morphologically defined MSC subpopulations. The use of single-cell optical imaging coupled with single-cell spatial lipidomics could assist in optimizing the MSC production process and improve mechanistic understanding of manufacturing process effects on MSC immune activity and heterogeneity.
间充质干细胞（MSC）疗法在临床试验中的效果有限，部分原因是免疫反应表型存在异质性。因此，在生物制造过程中需要技术来描述这些 MSC 的特性。已经发现细胞形态或形态与 MSC 的免疫反应性有关，但尚未建立单细胞形态和功能之间的直接关系。我们使用无标记的差分相差成像和基质辅助激光解吸/电离质谱成像（MALDI-MSI）来评估单细胞形态，并探索与脂质代谢免疫反应的关系。在干扰素γ（IFN-γ）刺激的 MSC 中，我们发现涉及细胞免疫功能的神经酰胺-1-磷酸（C1P）、磷脂酰胆碱（PC）、溶血磷脂酰胆碱（LysoPC）和甘油三酯（TAG）家族的脂质丰度更高。此外，我们还在形态定义的 MSC 亚群中鉴定出脂质特征的差异。单细胞光学成像与单细胞空间脂质组学的结合使用，可以辅助优化 MSC 的生产工艺，并深入了解制造工艺对 MSC 免疫活性和异质性的影响的机制。
