Extracellular vesicles derived from mesenchymal stem cells (MSCs) represent a novel approach for regenerative and immunosuppressive therapy. Recently, cytochalasin B-induced microvesicles (CIMVs) were shown to be effective drug delivery mediators. However, little is known about their immunological properties. We propose that the immunophenotype and molecular composition of these vesicles could contribute to the therapeutic efficacy of CIMVs. To address this issue, CIMVs were generated from murine MSC (CIMVs-MSCs) and their cytokine content and surface marker expression determined. For the first time, we show that CIMVs-MSCs retain parental MSCs phenotype (Sca-1+, CD49e+, CD44+, CD45−). Also, CIMVs-MSCs contained a cytokine repertoire reflective of the parental MSCs, including IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12(p40), IL-13, IL-17, CCL2, CCL3, CCL4, CCL5, CCL11, G-CSF, GM-CSF and TNF-α. Next, we evaluated the immune-modulating properties of CIMVs-MSCs in vivo using standard preclinical tests. MSCs and CIMVs-MSCs reduced serum levels of anti-sheep red blood cell antibody and have limited effects on neutrophil and peritoneal macrophage activity. We compared the immunomodulatory effect of MSCs, CIMVs and EVs. We observed no immunosuppression in mice pretreated with natural EVs, whereas MSCs and CIMVs-MSCs suppressed antibody production in vivo. Additionally, we have investigated the biodistribution of CIMVs-MSCs in vivo and demonstrated that CIMVs-MSCs localized in liver, lung, brain, heart, spleen and kidneys 48 h after intravenous injection and can be detected 14 days after subcutaneous and intramuscular injection. Collectively our data demonstrates immunomodulatory efficacy of CIMVs and supports their further preclinical testing as an effective therapeutic delivery modality.
The identification of rapid, reliable, and highly reproducible biological assays that can be standardized and routinely used in preclinical tests constitutes a promising approach to reducing drug discovery costs and time. This unit details a tandem, rapid, and reliable cell viability method for preliminary screening of chemical compounds. This assay measures metabolic activity and cell mass in the same cell sample using a dual resazurin/sulforhodamine B assay, eliminating the variation associated with cell seeding and excessive manipulations in assays that test different cell samples across plates. The procedure also reduces the amount of cells, test compound, and reagents required, as well as the time expended in conventional tests, thus resulting in a more confident prediction of toxic thresholds for the tested compounds. © 2016 by John Wiley & Sons, Inc.
The aim of the study – to accelerate the repair of the damaged brachial plexus using cells of the stromal vascular fraction isolated from adipose tissue.Materials and methods. The study was carried out in 62 patients using stromal-vascular fraction cells from adipose tissue and classical methods of treatment for brachial plexus injury. The effectiveness of regeneration was evaluated using electromyographic examination and positive recovery of motor and sensory function.Results and discussion. Assessment of the results of surgical treatment with stromal vascular fraction cells from adipose tissue after brachial plexus neurolysis revealed the restoration of early M3-M5 and S3-S4 functions in 90 % of patients, and in the comparison group – 68 % respectively. The number of patients with M4-M5 functions in the group using the stromal vascular fraction for brachial plexus neurotization was 85 %, while in the control group it was 64 %, respectively. Electroneuromyography data also indicated an increase in the average number of motor units by 30 % after using cells of the stromal-vascular fraction from adipose tissue, in contrast to the comparison group.Conclusion. Stromal vascular cells isolated from adipose tissue appear to be promising stimulants of brachial plexus injury repair.
The paper describes the results of clinical testing of an apparatus for dosed traction of brachial plexus trunks. It is shown that in the presence of a 5 cm diastasis, it is possible to connect the nerve fragments and perform neurorhaphy without tension by bringing the shoulder to the head and bring the shoulder to the physiological position two and a half months after the reconstructive surgery.
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