MSC-based cell therapy as a promising therapeutic strategy is used in various clinical fields, including tissue regeneration and immunomodulatory therapy.
MSCs are considered to be relatively safe in terms of genomic instability compared to induced pluripotential stem (iPS) cells and embryonic stem (ES) cells (Kim et al. , 2017). In the advancement of stem cell therapy, attempts are being made to develop an efficient production system to produce sufficient clinically relevant numbers of MSCs in a timely manner (Han et al. , 2014).
The aim of this study was to investigate of MRI effects on stemness genes (Oct-4, Sox-2 and Nanog) expressions on MSCs.
Our results are in consistent with previous studies that showed ectopic expression of stemness genes, such as Oct-4, Nanog, and Sox-2, promotes the proliferation potential of NIH 3T3 fibroblasts and MSCs while maintaining the lineage differentiation capacity (Go et al. , 2008, Zhang et al. , 2005).
Lengner and colleagues showed that oct-4 has no role in the self-renewal of somatic stem cell (Lengner et al. , 2007). Recent study revealed that Nanog maintains self-renewal capability of MSCs by delaying cellular senescence, increases the endogenous expression of Oct-4 and Sox-2, and preserves stemness (Park et al. , 2019).Many studies have shown that both Oct-4 and Sox-2 bind the Nanog promoter and induce Nanog expression (Park, Jun, 2019).Park et al. reported that secretome from Nanog-overexpressing MSCs accelerated the telogen-to-anagen transition in hair follicles and could be an excellent candidate as a powerful anagen inducer and hair growth stimulator for the treatment of alopecia (Park, Jun, 2019).
In this study, we also showed that long term effect of exposure to 1.5 T MRI for 10 min enhanced colony formation capacity (approximately 25 folds) of WJ-MSCs and increased number of active stem cells. In addition, the findings of the flow cytometry revealed that MRI treatment does not influence percentages of WJ-MSCs in apoptosis. Result showed that short term exposure to magnetic field did not induce apoptosis after 24 and 48 hours.
In the current investigation, we evaluated the cytotoxic effects of MRI treatment on WJ-MSCs using the MTT assay. MTT assay determines cell viability by quantitative assessment of a metabolic product and indicates activity of mitochondria in living cells; which has a direct relationship with cell proliferation and longevity (Tabatabaei et al. , 2015). The results indicated that MRI does not significantly impact on metabolic activity, mitochondria function and viability of WJ-MSCs.
In addition, we utilized doubling time assay to investigate the average duration of cell growth and division. The findings reported that MRI can decrease the average division time of WJ-MSC in short time, however, colony formation assay showed that this change was not permanent and reversed after a short time.
The result of MRI effect on regrowth of the cryopreserved MSCs suggests that the cell death of all MRI-treated MSCs may be due to increased sensitivity of these cells to the physical and chemical stress caused by the freezing process.
There is growing evidence that the increase in ROS content is the most common event for various types of cells after cryopreservation (Savitskaya and Onishchenko, 2016).
It has been shown that cryopreservation procedures can induce both apoptosis and necrosis (Savitskaya and Onishchenko, 2016). It was previously shown that during thawing in mesenchymal stem cells, apoptosis was induced by the activation of apoptosis-related proteins including Ca2+-dependent protease calpain, caspase-8, -9, and -3(Bissoyi and Pramanik, 2014).
Previous investigations have demonstrated that extended exposure to magnetic field decreased the viability percent and/or proliferation rate of stem cell and terminally differentiated somatic cells(Javani Jouni et al. , 2013, Marędziak et al. , 2014, Raylman et al. , 1996, Rosen and Chastney, 2009).
Seven-day exposure to 0.5 t static magnetic fields (SMF) inhibited viability, proliferation, cytokines secretion, surface antigen expression, and adipogenic and osteogenic differentiation of adipose-derived stem cells (ASCs) (Wang et al. , 2016). Several studies have demostrated that different intensities of the magnetic field and durations of exposure did not cause DNA damage in cells, such as ASCs (Wang, Xiang, 2016), THP1 (Amara et al. , 2007), peripheral blood mononuclear cells (Reddig et al. , 2015) and leukocytes(Kubinyi et al. , 2010).
Gruchlik et al. demonstrated that exposure to a SMF (300mT) for 24 hours inhibited the IL-6 secretion in normal human colon myofibroblasts (Gruchlik et al. , 2012).
Vergallo et al. reported that SMF exposure (1.4T, 24 h) had a significant inhibitory effect on the release of IL-6, IL-8, and INF-α from macrophage (Vergallo et al. , 2013).
In another study, exposure to a strong SMF (4.75T) for 24–48 hours significantly decreased the production of IL-2 in human peripheral blood mononuclear cells (Aldinucci et al. , 2003).
According to previous studies, it seems that the exposure to magnetic fields is associated with epigenetic changes but not genetic changes (mutations or DNA damage).
Review of the current literature suggests that Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) have the best potential to differentiate into adipocytes, chondrocytes, osteocytes, muscle cells, neurons, cardiomyocytes, and hepatocytes (Beeravolu et al. , 2017).
WJ-MSCs have been widely explored for cell-based therapy of immune-mediated, inflammatory, and degenerative diseases, due to their remarkable anti-inflammatory, immunosuppressive, immunomodulatory and regenerative potentials (Liau et al. , 2020, Noronha et al. , 2019).
Overall, our findings suggest that MRI treatment could be a useful approach to produce high-quality MSCs by improving their stemness properties.
This strategy has great potential for developing pluripotency in WJ-MSCs to become more versatile in clinical applications.
To the best of our knowledge, no other study has investigated the MRI effects on the stemness genes expression of MSCs.
However, it is worth noting that cancer stem cells, as a key driver of tumor formation and metastasis, express deregulated stemness-associated genes, such as OCT-4, Nanog, SOX-2 (Müller et al. , 2016). In addition, the personal exposure to MRI has the potential for deregulation of stemness related genes in normal stem cells. Therefore, it seems that precautions should be taken to reduce the possible risks of MRI exposure. Further studies are needed to determine MRI-treated MSCs have no adverse or unwanted effects following cell therapy. Our findings suggest that MRI treatment may be a useful strategy for gaining high-quality MSCs in clinical studies of MSC-based therapies.
Finally, preclinical and clinical studies should be designed to identify whether MRI treatment can be an effective approach to improve the therapeutic function of MSCs.