Labeling of human mesenchymal stem cells with different classes of vital stains: robustness and toxicity

• Published in: Stem cell research & therapy 2019-06, Vol.10 (1), p.187-187, Article 187
• Main Authors: Andrzejewska, Anna, Jablonska, Anna, Seta, Martyna, Dabrowska, Sylwia, Walczak, Piotr, Janowski, Miroslaw, Lukomska, Barbara
• Format: Article
• Language: English
• Subjects: Analysis; Cell labeling; Clinical trials; Diagnostic imaging; Epidermal growth factors; Fluorescent dyes; Gene expression; Genes; Genetic research; Health aspects; Labels; Medical research; Mesenchymal stem cells; Messenger RNA; MRI contrast agents; RNA; Stem cell transplantation; Stem cells; Surgery; Toxicity; Vital stains
• ISSN: 1757-6512; 1757-6512
• DOI: 10.1186/s13287-019-1296-8

Mesenchymal stem cell (MSC) transplantation has been explored as a new clinical approach to repair injured tissues. However, in order to evaluate the therapeutic activity of MSC, cell tracking techniques are required to determine the fate of transplanted cells in both preclinical and clinical studies. In these aspects, different vital stains offer the potential for labeling and monitoring of grafted cells in vivo. It is desirable to have tracking agents which have long-term stability, are not toxic to the cells, and do not affect cell function. Here, we selected three different labels: CellTracker™ Green CMFDA, eGFP-mRNA (genetic pre-tag), and Molday ION Rhodamine B™ (nanoparticle-based fluorescent and magnetic label) and performed extensive analysis of their influence on in vitro expansion of human bone marrow-derived mesenchymal stem cells (hBM-MSCs), as well as potential of affecting therapeutic activity and the impact on the durability of staining. Our study showed that basic hBM-MSC characteristics and functions might be affected by labeling. We observed strong alterations of metabolic activity and morphology after eGFP and CellTracker™ Green CMFDA hBM-MSC staining. Molday ION Rhodamine B™ labeling revealed superior properties relatively to other vital stains. The relative expression level of most of the investigated growth factors remained stable after cell labeling, but we have observed some changes in the case of EGF, GDNF, HGF, and IGF gene expression. Taken together, we suggest performing similar to ours extensive analysis prior to using any cell label to tag MSC in experiments, as it can thoroughly bias results.