Abstract
Over the past decade, cell therapy has found many applications in the treatment of different diseases. Some of the cells already used in clinical practice include stem cells and CAR-T cells. Compared with traditional drugs, living cells are much more complicated systems that must be strictly controlled to avoid undesirable migration, differentiation, or proliferation. One of the approaches used to prevent such side effects involves monitoring cell distribution in the human body by any noninvasive technique, such as magnetic resonance imaging (MRI). Long-term tracking of stem cells with artificial magnetic labels, such as magnetic nanoparticles, is quite problematic because such labels can affect the metabolic process and cell viability. Additionally, the concentration of exogenous labels will decrease during cell division, leading to a corresponding decrease in signal intensity. In the current work, we present a new type of genetically encoded label based on encapsulin from Myxococcus xanthus bacteria, stably expressed in human mesenchymal stem cells (MSCs) and coexpressed with ferroxidase as a cargo protein for nanoparticles’ synthesis inside encapsulin shells. mZip14 protein was expressed for the enhancement of iron transport into the cell. Together, these three proteins led to the synthesis of iron-containing nanoparticles in mesenchymal stem cells—without affecting cell viability—and increased contrast properties of MSCs in MRI.
Highlights
While using cells for therapy or establishing preclinical models, it is critically important to maintain the possibility of long-term cell tracking after administration into the organism
Luciferase (Fluc), Renilla luciferase (Rluc), and bacterial luciferases [8] have been utilized for this purpose
We present genetically encoded labels based on the Myxococcus xanthus (Mx) encapsulin system expressed in human adipose-derived mesenchymal stem cells
Summary
While using cells for therapy or establishing preclinical models, it is critically important to maintain the possibility of long-term cell tracking after administration into the organism. The labels used for such long-term tracking should have minimal effects on the processes of cell division, migration, and proliferation—and should not change cell viability. The main disadvantage of these labels is their limited depth of detection (usually not more than 2 mm); this approach is only possible in surface tissues of small animals such as mice. Bioluminescent imaging is another viable alternative based on the reaction between the enzyme luciferase and its substrate luciferin. The bioluminescent approach is more sensitive than the fluorescent approach, but the depth of signal penetration through the tissues is still limited to a few centimeters
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