Vladimir Zelman Center for Neurobiology and Brain Rehabilitation, Skolkovo Institute of Science and Technology, Skolkovo Institute of Science and Technology, Bolshoy Boulevard 30, 121205 Moscow, Russia.
Science Medical Center, Saratov State University, 83 Astrakhanskaya Str., 410012 Saratov, Russia.
Int J Mol Sci. 2023 Sep 4;24(17):13665. doi: 10.3390/ijms241713665.
The behavior and migration of human mesenchymal stromal cells (hMSCs) are focal points of research in the biomedical field. One of the major aspects is potential therapy using hMCS, but at present, the safety of their use is still controversial owing to limited data on changes that occur with hMSCs in the long term. Fluorescent photoconvertible proteins are intensively used today as "gold standard" to mark the individual cells and study single-cell interactions, migration processes, and the formation of pure lines. A crucial disadvantage of this method is the need for genetic modification of the primary culture, which casts doubt on the possibility of exploring the resulting clones in personalized medicine. Here we present a new approach for labeling and tracking hMSCs without genetic modification based on the application of cell-internalizable photoconvertible polyelectrolyte microcapsules (size: 2.6 ± 0.5 μm). These capsules were loaded with rhodamine B, and after thermal treatment, exhibited fluorescent photoconversion properties. Photoconvertible capsules demonstrated low cytotoxicity, did not affect the immunophenotype of the hMSCs, and maintained a high level of fluorescent signal for at least seven days. The developed approach was tested for cell tracking for four days and made it possible to trace the destiny of daughter cells without the need for additional labeling.
人骨髓间充质干细胞（hMSCs）的行为和迁移是生物医学领域研究的重点。其中一个主要方面是利用 hMSC 进行潜在治疗，但由于长期以来 hMSC 发生变化的数据有限，其使用的安全性仍存在争议。荧光光转化蛋白目前被广泛用作标记单个细胞、研究单细胞相互作用、迁移过程和形成纯系的“金标准”。该方法的一个关键缺点是需要对原代培养进行基因修饰，这使人对探索个性化医学中产生的克隆的可能性产生了怀疑。在这里，我们提出了一种新的方法，无需基因修饰即可对 hMSC 进行标记和跟踪，该方法基于应用可内化细胞的光转化聚电解质微胶囊（大小：2.6 ± 0.5 μm）。这些胶囊中装载了罗丹明 B，经过热处理后，表现出荧光光转化特性。光转化胶囊表现出低细胞毒性，不影响 hMSC 的免疫表型，并能保持至少七天的高荧光信号水平。该方法已在细胞跟踪测试中进行了四天，无需额外标记即可追踪子细胞的命运。
