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

Abstract

Background 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. Methods 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. Results 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. Conclusions 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. Electronic supplementary material The online version of this article (10.1186/s13287-019-1296-8) contains supplementary material, which is available to authorized users.

Neuro‐Cells therapy improves motor outcomes and suppresses inflammation during experimental syndrome of amyotrophic lateral sclerosis in mice

“…infusion of a preparation of nonmanipulated human stem cells “Neuro‐Cells” obtained from bone marrow, which are a combination of both mesenchymal stem cells (MSCs) and hemopoietic stem cells (HSCs). As both MSCs and HSCs have been shown to have the ability to differentiate into a spectrum of adult cell populations, many studies have sought to examine either the combined or individual contributions to repair in vivo in models of injury and potentially capitalize on the relationship between the two cell populations that is known to exist . As for instance, MSCs were shown to act as a feeder layer maintaining HSCs in an undifferentiated state.…”

Mesenchymal Stem Cells Do Not Lose Direct Labels Including Iron Oxide Nanoparticles and DFO-89Zr Chelates through Secretion of Extracellular Vesicles

“…Moreover, we have shown that magnetic labeling of MSCs does not affect EV production in terms of the number of EVs, their size, as well as the presence of tetraspanins on their surface, which is also very positive information for the potential of cellular labeling and therapeutic transplantation of transplanted cells. This is important, as we have previously demonstrated that SPION has negligible impact on the MSCs’ stemness characteristic and their other properties in vitro [ 14 ]. Therefore, it seems that direct labels are quite resistant to packaging them to the EVs, so there is minimal chance that cell labeling will diminish EV-mediated positive therapeutic consequences of MSC transplantations.…”

Mesenchymal Stem Cells for Neurological Disorders

“…Superparamagnetic iron oxide nanoparticles (SPIONs) are quite versatile and relatively frequently used cellular label. It has been shown that SPIONs are mostly neutral to MSCs in vitro [ 318 ] and in vivo. [ 319 ] A very high signal of SPIONs, which allows them to be detected within seconds, is a major advantage.…”