The review considers the use of mesenchymal stem cells (MSCs) derived from various tissues in treating the conditions consequent to ischemia (circulation injury) and subsequent reperfusion (blood supply restoration) of organs and tissues. MSC modification methods to improve the treatment efficacy are described. A potential effect in treating frostbites is hypothesized for MSCs.

Frostbite is the most common type of traumatic injuries in the cold climate of the Arctic. Post-traumatic recovery of the patients takes a long time and often leads to the loss of the damaged limb. A promising approach for frostbite therapy could be treatment with mesenchymal stem/stromal cells (MSCs), which are widely used in regenerative medicine for different applications. Here, we aimed to discuss the possibility of frostbite treatment with MSCs. The success of MSC use for therapy of different injures depends on multiple factors including the source and functional state of MSCs. These aspects along with strategies for improvement of therapeutic effect of MSC treatment are also discussed in detail.

The use of mesenchymal stem cells is an effective treatment strategy for a number of retinal degenerative diseases. The poor survival of these cells after transplantation is the limiting factor of this approach. It was shown previously that the cultivation of mesenchymal stem cells under hypoxia can increase their proliferative activity. We assumed that this method of cultivation will improve the viability of these cells after their transplantation into the subretinal space. To this end, we isolated mesenchymal stem cells from the red bone marrow of mice, characterized their phenotype, their ability to differentiate in the chondrogenic, osteogenic, and adipogenic directions, as well as their proliferative activity after cultivation under hypoxia (5% oxygen in the atmosphere) and normoxia (21% oxygen in the atmosphere). Red bone marrow cells were obtained in the same way from C57 Black mice carrying the GFP gene. These cells were loaded with magnetic microparticles after preliminary cultivation under normoxia (the control) and hypoxia (the experiment) and injected subretinally to rabbits. The cells were held at the injection site using a magnetic implant that prevented their migration. Cell survival was evaluated on the 3rd, 5th, 9th, 12th, and 15th days using fluorescence microscopy and optical coherence tomography. According to the obtained data, the cells grown under hypoxic conditions remained viable in the subretinal space for 9 days, while the cells that grew under normoxia conditions died after 6 days. Thus, pre-cultivation of mesenchymal stem cells under hypoxic conditions can increase their viability after transplantation into the subretinal space, which can be used in the treatment of degenerative diseases of the retina.

Quantitative determination of cytokines, chemokines, growth factors, and other soluble protein molecules in various biological fluids is a routine task of modern diagnostics and biomedicine; however, currently existing methods of analysis include several stages and take a long time. This paper presents a protocol that allows determination of an analyte concentration in a sample within a few minutes using biomolecule layer interferometry. Conditioned media in which mouse mesenchymal stem cells were grown under normoxia and hypoxia in the form of monolayer, 3D cell sheets, or 3D spheroids were used as a model system, in which the concentration of VEGF-A (vascular endothelial growth factor) was determined. The method showed high sensitivity (at least 0.10 ng/mL of VEGF-A). The obtained values of VEGF-A concentrations in mesenchymal stem cell conditioned media were close to the values determined by enzyme immunoassay. Using biomolecule layer interferometry, a significant 2.5–3.0 fold increase in VEGF-A production by spheroids and cell sheets of mesenchymal stem cells was found in comparison with the cells grown in a monolayer. Thus, the developed protocol made it possible to adapt the method of biomolecule layer interferometry for rapid quantitative analysis of growth factors in the conditioned media.

Quantitative analysis of cytokines, chemokines, growth factors and other soluble proteins in different biological liquids is routinely performed in contemporary diagnostics and biomedicine research. However, current methods of analysis are time-consuming and include multiple steps. In this study, we have developed a protocol that describes how bio-layer interferometry can be applied to quantify an analyte in several minutes. Conditioned growth media collected from mouse mesenchymal stem cells grown in normoxia or hypoxic conditions in a monolayer fashion, MSC-derived 3D cell sheets and 3D spheroids were used as a model system in which we determined a concentration of vascular endothelial growth factor (VEGF-A). This technique displayed a high sensitivity (down to 0.1 ng of VEGF-A per mL as a minimum). The measured concentrations of VEGF-A in the conditioned media from mesenchymal stem cells turned out to be similar with values determined by the enzyme-linked immunosorbent assay. Using bio-layer interferometry, it was shown that as compared to mesenchymal stem cells grown in monolayer, spheroids and 3D sheets of mesenchymal stem cells produce significantly more VEGF-A (by 2.5-3.0-fold). Thus, due to the developed protocol it was possible to adapt bio-layer interferometry for rapid quantification of growth factors in conditioned media.

The use of mesenchymal stem cells is an effective treatment strategy for a number of retinal degenerative diseases. Limited survival of these cells following transplantation remains a significant barrier in achieving a therapeutic effect. It has been shown earlier that hypoxia-grown mesenchymal stem cells display enhanced proliferative activity. We hypothesized that such cultivation condition could be beneficial for successful injection of these cells into the subretinal region. For this purpose, we isolated mesenchymal stem cells from mouse red bone marrow, described phenotypic characterization and the ability of these cells to chondrogenic, osteogenic and adipogenic differentiation, and proliferative activity in hypoxic (5% of oxygen in the atmosphere) and normoxic (21% of oxygen in the atmosphere) conditions. In the same manner, we isolated mesenchymal stem cells from red bone marrow obtained from GFP-expressing C57 black mice. After preliminary cultivation under normoxia (control cells) or hypoxia (experimental cells), the cells were loaded with magnetic particles and injected into the subretinal region of rabbit eyes...

The purpose of this review is to systematize scientific data and to assess the prospects for using mesenchymal stem cells (MSCs) to treat injury in the Arctic. Here, we considered possible sources of MSCs, barriers and limitations of their use and possible strategies to improve their therapeutic outcome (bibliography: 33 refs).

Целями данного обзора являются систематизация научных данных и оценка перспектив использования мезенхимальных стволовых клеток для лечения травм различного генеза в условиях Арктики. Рассмотрены возможные источники мезенхимальных стволовых клеток, барьеры и ограничения, связанные с их использованием, и стратегии для улучшения их терапевтических свойств (библ.: 33 ист.).

The effect of different polyoxyethylene–polyoxypropylene triblock copolymers, their concentration, and mode of action on the loading of poly-(lactic-co-glycolic acid) copolymer-based microparticles containing such medicinal agents as radachlorin (chlorin e 6 ) or ethidium bromide in mesenchymal stem cells was studied. It has been shown that medicinal agents encapsulated inside microparticles affect the loading of these particles in the cytoplasm of mesenchymal stem cells. The number of cells that absorbed the particles with chlorin e 6 is approximately two times lower than that in the experiments with ethidium bromide. It has been shown that pretreatment of microparticles with triblock copolymers is more efficient for loading them in cells compared with simultaneous introduction of triblock copolymers and particles into the culture medium. Treatment of ethidium bromide-containing microparticles with triblock copolymers is not efficient for their loading in mesenchymal stem cells compared to the control. The exception is Pluronic 123; when particles are treated with it at concentrations of 1 and 2%, the loading of particles in cells increases compared to the control by factors of approximately 11 and 5, respectively. For particles with chlorin e 6 , their pretreatment with triblock copolymers at a concentration of 4% is most efficient; the loading of the pretreated particles in cells is increased by factors of approximately 3 to 11.

The article presents statistical data on the time characteristics of petit mal epilepsy registered in the course of prolonged telemetric EEG recording in 3 epileptic patients in the process of treatment. The time-course of the frequency and also the average and total daily length of attacks during sleep or wakefulness are analyzed. The pattern of the genesis of paroxysms was found to be different: paroxysms were more likely both to arise and stop in sleep than in wakefulness. Nocturnal attacks were noted to be more resistant to therapeutic measures as compared with diurnal ones.