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AIM: To evaluate the efficacy of transplantation of human umbilical cord mesenchymal stem cells(hUCMSCs)in the treatment of corneal alkali burn in rabbits, and study the infiltration of polymorphonuclear neutrophils(PMNs)and the changes of vascular endothelial growth factor(VEGF)expression.METHODS: Corneal alkali burn models were established in right eyes of 75 healthy Japanese white rabbits, which were divided into three groups(group A, B and C), with 25 rabbits in each group. Group A was treated with amniotic membrane combined with hUCMSCs on the day after corneal alkali burn. Group B was treated with amniotic membrane only. Group C did not give any treatment after corneal alkali burn. At 3, 7, 14, 21 and 28d after corneal alkali burn, the corneal recovery was observed by slit lamp and photographed, the growth of corneal neovascularization(CNV)was scored, and corneal tissue was separated to make pathological sections. PMNs infiltration was observed by hematoxylin-eosin(HE)staining, and the expression of VEGF was determined by immunohistochemical staining.RESULTS: The growth of CNV in group A was much slower than that in group B at 14d after alkali burn. The CNV growth score around lesions of group A was significantly lower than that of group B(P<0.05). The quantity of PMNs increased on the 3d with the stromal layer of cornea infiltrated, relatively decreased on the 7d, shown a peak on the 14d, and then decreased gradually. Early infiltration after alkali burn was in the corneal stroma of the lesion area, and the extent of infiltration was equal to the ulcer area at later stage. The cell densities of corneal PMNs in group A and group B were significantly lower than those in group C at all time points after alkali burns(P<0.05), and those in group A were significantly lower than group B at 14 and 21d(P<0.05). The expression levels of corneal VEGF in all groups after alkali burn reached peak at 7～14d and decreased significantly at 28d, and the expression levels of VEGF in group A and group B at all time points after alkali burn were significantly lower than those in group C(P<0.05), and group A was significantly lower than that in group B at 7, 14 and 21d(P<0.05).CONCLUSION: The transplantation of hUCMSCs after alkali burn cornea can reduce the formation of CNV and inhibit corneal revascularization after alkali burn. The corneal pathological lesions and vascularization are closely related to PMNs and VEGF.
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Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease affecting both upper and lower motor neurons in the brain and spinal cord. One important aspect of ALS pathogenesis is superoxide dismutase 1 (SOD1) mutant-mediated mitochondrial toxicity, leading to apoptosis in neurons. This study aimed to evaluate the neural protective synergistic effects of ginsenosides Rg1 (G-Rg1) and conditioned medium (CM) on a mutational SOD1 cell model, and to explore the underlying mechanisms. We found that the contents of nerve growth factor, glial cell line-derived neurotrophic factor, and brain-derived neurotrophic factor significantly increased in CM after human umbilical cord mesenchymal stem cells (hUCMSCs) were exposed to neuron differentiation reagents for seven days. CM or G-Rg1 decreased the apoptotic rate of SOD1G93A-NSC34 cells to a certain extent, but their combination brought about the least apoptosis, compared with CM or G-Rg1 alone. Further research showed that the anti-apoptotic protein Bcl-2 was upregulated in all the treatment groups. Proteins associated with mitochondrial apoptotic pathways, such as Bax, caspase 9 (Cas-9), and cytochrome c (Cyt c), were downregulated. Furthermore, CM or G-Rg1 also inhibited the activation of the nuclear factor-kappa B (NF-κB) signaling pathway by reducing the phosphorylation of p65 and IκBα. CM/G-Rg1 or their combination also reduced the apoptotic rate induced by betulinic acid (BetA), an agonist of the NF-κB signaling pathway. In summary, the combination of CM and G-Rg1 effectively reduced the apoptosis of SOD1G93A-NSC34 cells through suppressing the NF-κB/Bcl-2 signaling pathway (Fig. 1 is a graphical representation of the abstract).
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Humans , NF-kappa B/metabolism , Ginsenosides/pharmacology , Amyotrophic Lateral Sclerosis/genetics , Culture Media, Conditioned/pharmacology , Superoxide Dismutase-1 , Neurodegenerative Diseases , Neurons/metabolism , ApoptosisABSTRACT
ObjectiveThis study aimed to investigate the effects of eugenol on inhibiting the inflammatory activation of human umbilical cord mesenchymal stem cells （HUC-MSCs） and the pro-inflammatory phenotype of hepatic stellate cells （HSCs） in liver fibrosis， and to explore their underlying mechanisms. MethodsHUC-MSCs were cultured and identified in vitro， and the toxicity of eugenol to HUC-MSCs was evaluated by MTT method. The effect of eugenol on the migration ability of HUC-MSCs was investigated by in vitro scratch test. The expression of α-SMA， COL1A1， Smad2/3 and p-Smad2/3 of LX-2 cells activated by TGF-β1 treated with EU-MSCs-CM and MSCs-CM were detected by WB assay. EU-MSCs-CM and MSCs-CM treated THP-1 macrophages stimulated with Lipopolysaccharide （LPS） were analyzed for the expression of surface markers CD11b， CD86， and CD206 by flow cytometry. Additionally， the expression of pro-inflammatory genes TNF-α， IL-1β， and IL-6 in THP-1 macrophages was detected by qPCR. ResultsThe results of MTT method showed that the viability of the cells remained above 90% after 24 h and 48 h treatment at 0， 7.5， 15 μg/mL. In vitro scratches showed that eugenol treatment enhanced HUC-MSCs migration. WB results showed that compared with MSCs-CM treatment， EU-MSCs-CM treatment significantly inhibited the expression of α-SMA， COL1A1， Smad2/3， and p-Smad2/3 of activated HSCs. Flow cytometry showed that compared with MSCs-CM treatment， EU-MSCs-CM treatment had a more significant inhibitory effect on CD86， a M1-type polarization marker in THP-1 macrophages. The results of qPCR experiment showed that compared with MSCs-CM treatment， EU-MSCs-CM treatment more significantly inhibited the expressions of TNF-α， IL-1β and IL-6 of THP-1 macrophage proinflammatory genes. ConclusionsEugenol enhances the inhibitory effect of HUC-MSCs on inflammatory activation of HSCs， possibly by regulating TGF-β1/Smads signaling pathway. It also enhances the inhibitory effect of HUC-MSCs on the pro-inflammatory phenotype of macrophages. Proinflammatory macrophages can promote inflammatory activation of HSCs.
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Impaired immunomodulatory capacity and oxidative stress are the key factors limiting the effectiveness of mesenchymal stem cell transplantation therapy. The present study was aimed to investigate the effects of jujuboside A (JuA) on the protective effect and immunomodulatory capacity of human umbilical cord mesenchymal stem cells (hUC-MSCs). Hydrogen peroxide was used to establish an oxidative damage model of hUC-MSCs, while PBMCs isolated from rats were used to evaluate the effect of JuA pre-treatment on the immunomodulatory capacity of hUC-MSCs. Furthermore, Hoechst 33258 staining, lactate dehydrogenase test, measurement of malondialdehyde, Western blot, high-performance liquid chromatography; and flow cytometry were performed. Our results indicated that JuA (25 μmol·L-1) promoted the proliferation of hUC-MSCs, but did not affect the differentiating capability of these cells. JuA pre-treatment inhibited apoptosis, prevented oxidative damage, and up-regulated the protein expression of nuclear factor-erythroid factor 2-related factor 2 and heme oxygenase 1 in hUC-MSCs in which oxidative stress was induced with H2O2. In addition, JuA pre-treatment enhanced the inhibitory effect of hUC-MSCs against abnormally activated PBMCs, which was related to stimulation of the expression and activity of indoleamine 2,3-dioxygenase. In conclusion, our results demonstrate that JuA pre-treatment can enhance the survival and immunomodulatory ability through pathways related to oxidative stress, providing a new option for the improvement of hUC-MSCs in the clinical setting.
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Animals , Humans , Rats , Cell Differentiation , Hydrogen Peroxide/metabolism , Mesenchymal Stem Cells , Oxidative Stress , Saponins , Umbilical Cord/metabolismABSTRACT
Objective:To explore the effect of photobiomodulation (PBM) combined with umbilical cord mesenchymal stem cell transplantation on motor function recovery in rats with spinal cord injury.Methods:The mesenchymal stem cells were irradiated with a laser energy density of 1.5, 3, 6 and 12 J/cm 2. The optimal energy density was screened by the MTT method on the 3rd day. Before cell transplantation, 32 male SD rats were randomly divided into the spinal cord injury group, which was injected with normal saline without cells; the stem cell transplantation group, which was injected with stem cells in the injury model; the laser irradiation group, which was injected with cell-free saline and laser irradiation; and the combined treatment group, which was treated with cell transplantation and laser irradiation. BBB score and inclined plate test were performed on the 1st, 3rd, 7th, 14th and 21st day, and hematoxylin-eosin staining and Nissl staining were performed on the 21st day. Results:The laser irradiation with an energy density of 12 J/cm 2 can accelerate cell proliferation ( P<0.05). After the modeling, the BBB score of the combined treatment group was higher than that of the other groups (all P<0.05), and the motor function recovered significantly. In the inclined plate experiment, the performance of the combined treatment group and the laser irradiation group was also better than that of other groups. The results of hematoxylin-eosin staining showed that the cavity area in the combined treatment group was significantly reduced, and the inflammatory reaction was the lightest. The staining of Nissl bodies became deeper, and the spinal cord injury was significantly reduced. Conclusions:PBM can promote the recovery of motor function in rats with spinal cord injury after the transplantation of umbilical cord mesenchymal stem cells, and has an obvious therapeutic effect on spinal cord injury in rats. This study provides a basis for the treatment of spinal cord injury.
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Objective:To establish a three-level clinical grade human umbilical cord mesenchymal stem cells (hUC-MSCs) bank, including seed cell bank (SCB), master cell bank (MCB) and working cell bank (WCB), and provide hUC-MSCs with controllable quality for clinical research and application.Methods:247 human umbilical cord tissues were isolated, cultured, amplified, subcultured and frozen in GMP laboratory, and the biological characteristics, safety and stability of hUC-MSCs were tested in accordance with the requirements of relevant quality management control specifications.Results:247 strains of hUC-MSCs were isolated and prepared. The prepared hUC-MSCs have good purity and homogeneity without tumorigenicity, show good differentiation ability in biological efficacy, and have strong immunosuppressive effect in the process of co-culture with immune cells. These cells have passed the quality check of National Institute for Food and Drug Control. In this study, a three-level hUC-MSCs bank was established, and it was included into the National Stem Cell Translational Resource Center.Conclusions:A three-level clinical hUC-MSCs bank was successfully established and preliminarily applied to clinical research, which effectively promoted the standardized development of clinical stem cell resource bank and the clinical transformation and application of stem cells in China.
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Objective To study the effect of human umbilical cord mesenchymal stem cells (hUCMSCs) tail vein transplantation on depressive behavior in chronic unpredictable mild stress (CUMS) model mice. Methods The third generation hUCMSCs were abtained; Sixty C57BL/6 male mice were randomly divided into the normal model group (+ normal saline), the cell group (+hUCMSCs) and the fluoxetine(flu) group (+flu), with 15 mice in each group. The depression model of CUMS mice with a 6 week duration was constructed; From the third week, hUCMSCs and normal saline were transplanted into the mice by tail vein injection, and the mice were gavaged with flu daily from the fifth week; Weight changes in each group were recorded weekly; The depression model and therapeutic effect of mice were evaluated by sucrose preference test, tail suspension test, forced swimming test and open field test; The cell changes of CAI and CA3 region in hippocampus were observed by HE staining; Transmission electron microscopy was used to detect the changes of synapses in CA3 region of hippocampus; Real-time PCR was used to detect mRNA expressions of synaphysin (SYP) and postsynaptic density protein 95 (PSD-95), which are key proteins of synaptic plasticity. Results hUCMSCs improved weight decrease in depressed mice; behavioral experiments observed that the model group mice showed depressive behavior, while the cell group and the drug group mice depression were improved; HE staining showed that, compared with the model group, cells in CA 1 and CA3 regions of the hippocampus of mice in the cell group and the group were arranged orderly and the number of cells increased; Transmission electron microscopy showed that compared with the model group, the number of synapses in the CA3 region of the hippocampus was more, the synaptic gap was narrower, and the density of synaptic spines was higher in the cell group and the drug group; More mRNA expressions of SYP and PSD-95 were detected by Real-time PCR in the cell group and the drug group than in the model group. Conclusion hUCMSCs transplatation showes antidepressant effects, which are associated with improved synaptic plasticity in the hippocampus.
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BACKGROUND: Stem cell transplantation has a significant neuroprotective effect on neurological diseases. Current transplantation methods such as arteriovenous transplantation and brain stereotactic transplantation are not suitable for clinical application in preterm infants. OBJECTIVE: To explore the feasibility of nasal transplantation of human umbilical cord mesenchymal stem cells and human neural stem cells for the treatment of white matter injury in premature rat infants. METHODS: Human umbilical cord mesenchymal stem cells were prepared from human umbilical cord tissue, and human neural stem cells were prepared from human embryonic brain tissue. In vitro migration of two kinds of cells was assessed by Transwell method. Forty 3-day-old Sprague-Dawley rats were randomly divided into sham operation group, model control group, human umbilical cord mesenchymal stem cell transplantation group and human neural stem cell transplantation group, with 10 rats in each group. Rats in all groups except the sham operation group were treated with right common carotid artery ligation and hypoxia for 90 minutes to establish a rat model of white matter injury in the preterm infant. Totally 1×106 cells were delivered intranasally in the transplantation group at 3 days after injury. Each nostril was infused with 5×105, and each nostril was infused once. On day 7 after injury, MBP immunofluorescence staining was used to detect the expression of myelin basic protein in the white matter of the brain to identify the damage of the white matter injury model. At 24 hours after transplantation, human umbilical cord mesenchymal stem cell migration was detected by anti-HuNu immunohistochemical method and human neural stem cell migration was detected by CM-Dil fluorescent labeling method. RESULTS AND CONCLUSION: (1) On day 7 after modeling, compared with the normal side, the positive area of MBP decreased in cingulate band, corpus callosum and external capsule of the affected side in the model of brain white matter injury in preterm infants (P < 0.05), indicating a successful modeling. (2) In vitro experiments showed that the migration rate of human neural stem cells was the same as that of human umbilical cord mesenchymal stem cells. (3) At 24 hours after the nasal transplantation, human umbilical cord mesenchymal stem cells migrated to the cortex, corpus callosum and hippocampus on the normal side and the damaged side, and human neural stem cells migrated to the damaged cortex, corpus callosum and hippocampus, and human umbilical cord mesenchymal stem cells migrated more than human neural stem cells. (4) Overall, these findings indicate that 24 hours after the nasal transplantation, human umbilical cord mesenchymal stem cells could survive and migrate to the normal side and the injury side, and human neural stem cells could survive and migrate to the injury side; and the migration of human umbilical cord mesenchymal stem cells was more extensive than that of human neural stem cells.
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BACKGROUND: Systemic lupus erythematosus is an autoimmune disease with unknown causes. To establish a tree shrew model of systemic lupus erythematosus is helpful to understand its pathogenesis and provide evidence for stem cell transplantation in the treatment of autoimmune diseases. OBJECTIVE: To establish a tree shrew model of systemic lupus erythematosus and to assess the therapeutic effect of umbilical cord mesenchymal stem cell transplantation. METHODS: Tree shrews were grouped and intraperitoneally injected pristane, lipopolysaccharide, and their combination. At 3 weeks after injection, 12 tree shrew models were divided into treatment group and model control group (n=6 per group). An additional 6 models were selected as a normal control group. In the treatment group, each tree shrew was injected with 1×106 DiR-labeled umbilical cord mesenchymal stem cells through caudal vein. Two weeks later, the heart, liver, spleen, lung and kidney of tree shrews were taken for pathological sections. The sections received hematoxylin-eosin staining, kidney Masson staining and immune complex detection. Simultaneously, the heart, liver, spleen, lung and kidney of the three groups of tree shrews were taken for in vitro imaging. RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining showed pathological changes of the heart, liver, spleen, lung and kidney in the model control group; and there were a lot of immune complex deposits in renal tissue in the model control group. The pathological changes in the treatment group improved, and the structure recovered to close to the normal control group. (2) In vitro imaging showed that DiR-labeled cells were mainly distributed in the lung, liver and spleen of the tree shrews in the treatment group. The fluorescence intensity of tree shrews in the treatment group was significantly higher than that in the normal control group and model control group (P < 0.05). (3) Results demonstrated that intraperitoneal injection of pristane and lipopolysaccharide is the best method to induce pathological changes of systemic lupus erythematosus in tree shrews. The pathological changes after treatment with umbilical cord mesenchymal stem cells have improved, indicating that umbilical cord mesenchymal stem cells have certain treatment effects on tree shrew models of systemic lupus erythematosus.
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BACKGROUND: Transplanting islet-like cells induced by human umbilical cord mesenchymal stem cells (hUC-MSCs) into type 1 diabetic mice can reduce blood glucose level and improve the symptoms of diabetes mellitus. However, there are few reports on intraperitoneal transplantation. OBJECTIVE: To study the therapeutic effect of transplantation of islet-like cells induced by hUC-MSCs in different ways for the treatment of type 1 diabetic mice. METHODS: The hUC-MSCs were isolated and cultured by tissue explants adherent method and differentiated into islet-like cells. The 3 of 15 male C57BL/6J mice were used as normal group, and the remaining mice were taken to prepare a mouse model of type 1 diabetes using intraperitoneal injection of streptozotocin. After successful modeling, nine model mice were randomly divided into diabetes group, tail vein-islet-like cells group, and abdomen-islet-like cells group, with three mice in each group. After 10 days of modeling, the normal group and diabetic group were not treated. The tail vein-islet-like cells group was injected with 5×105 cells/0.4 mL islet-like cells via the tail vein and the abdomen-islet-like cells group was intraperitoneally injected with 5×105 cells/0.4 mL islet-like cells. During the treatment, the blood glucose and insulin levels were measured twice a week; glucose tolerance test was performed at 28 days after cell transplantation; and fasting insulin level was detected at 42 days after cell transplantation. RESULTS AND CONCLUSION: (1) Compared with the diabetic group, in the tail vein-islet-like cells group, the blood glucose level began to decrease on the 10th day after transplantation and maintained until the 31st day, and the insulin level and glucose tolerance significantly improved (P < 0.05). However, there was no significant improvement in blood glucose level, insulin level and glucose tolerance in the abdomen-islet-like cells group. (2) To conclude, transplantation of hUC-MSCs induced islet-like cells for the treatment of type 1 diabetic mice via tail vein is an ideal transplantation method, and the effect of intraperitoneal injection is unsatisfactory.
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BACKGROUND: The traditional treatment methods for ischemic heart disease can temporarily relieve pain and improve the quality of life, but cannot restore and regenerate the myocardial tissue. Stem cells have been reported to possess the potential functions of tissue regeneration and multiplex differentiation characteristics which provide a new opportunity for the ischemic heart disease treatment. Because of the unique advantages of umbilical cord mesenchymal stem cells including easy separation, no immune rejection reaction, no ethical dilemmas and remarkable immunomodulatory effect, umbilical cord mesenchymal stem cells have been considered as one of the most ideal candidate seeds for the treatment of ischemic heart disease. OBJECTIVE: To investigate the clinical efficacy of umbilical cord mesenchymal stem cells transplantation in patients with ischemic heart disease in 3-year followup. METHODS: From January 2013 to June 2016, eight patients with coronary heart disease were admitted to the First Hospital of Hebei Medical University and randomly divided into stem cell transplantation group and control group. Four patients in the stem cell transplantation group received conventional treatment and intravenous infusion of umbilical cord mesenchymal stem cells. Four patients in the control group only received conventional treatment. At 3 years after treatment, cardiac function indexes, biochemical indexes and ST segment changes of electrocardiogram lead II were evaluated in two groups. All patients signed the informed consent. The experiment was approved by the Ethics Committee of First Hospital of Hebei Medical University. RESULTS AND CONCLUSION: Patients in each group were alive after 3 years. Compared with the control group, left ventricular ejection fraction and left ventricular fraction shortening displayed an increasing trend in the stem cell transplantation group (P > 0.05). The changes of left ventricular ejection fraction and left ventricular fraction shortening indicated an important statistical improvement in the stem cell transplantation group compared with the control group (P 0.05). ST segment changes of different electrocardiogram leads were various before and after treatment in both groups. This study can only show that human umbilical cord mesenchymal stem cells can improve the cardiac function of patients with ischemic heart disease to a certain extent, but there is no significant improvement in the classification of cardiac function, biochemical indicators, and the influence on electrocardiogram leads is not clear. Thus, the significance of this treatment method needs further study.
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BACKGROUND: Vascular injury is a common complication after balloon dilatation. The development of umbilical cord mesenchymal stem cells (UC-MSCs) provides a new method for treating vascular injury. OBJECTIVE: To investigate the mechanism underlying the repair of damaged blood vessels by human UC-MSCs (hUC-MSCs) transfected with interleukin-8RA/B (IL-8RA/B) adenovirus. METHODS: hUC-MSCs and human umbilical vein endothelial cells (hUVECs) were collected and transfected with adenovirus vectors containing human IL-8RA and/or IL-8RB cDNAs and green fluorescent protein. A rat model of carotid artery injury was established. Sprague-Dawley rats were randomly divided into four groups: IL-8RA/B-hUCMSCs group, Il-8ra/B-hUVECs group, Null-hUCMSCs group, and control group, followed by injection of 0.5×106 corresponding cells (500 μL) and same volume of normal saline via the tail vein respectively at 1, 3, and 5 hours post-surgery. After 30 minutes of injection, the carotid artery was taken and the expression of green fluorescent protein was observed. After 24 hours, the serum levels of inflammatory and anti-inflammatory factors were measured by ELISA; and the infiltration of neutrophil cells and mononuclear macrophages was observed by immunohistochemistry. After 14 days, Evans blue staining was used to observe vascular endothelialization and fibrosis. After 28 days, the neointimal hyperplasia was observed by hematoxylin-eosin staining. RESULTS AND CONCLUSION: (1) After 30 minutes of IL-8RA/B-hUC-MSCs infusion, the expression of green fluorescent protein was observed in the injured vascular intima, and the fluorescence expression was higher than that of the other three groups. (2) After 24 hours of IL-8RA/B-hUC-MSCs infusion, the expression of inflammatory factors in the serum was significantly lower than that of the other three groups, while the expression of anti-inflammatory factor interleukin-10 was higher than that of the other three groups (P < 0.05). In addition, inflammatory cell infiltration in the IL-8RA/B-hUC-MSCs group decreased significantly. (3) hUC-MSCs overexpressing interleukin-8 receptor promoted re-endothelialization of injured vessels and reduced vascular fibrosis after 14 days of infusion. (4) IL-8RA/B-hUC-MSCs reduced vascular neointimal hyperplasia after 28 days of infusion. (5) Interleukin-8 receptor enhances the targeted homing ability of hUC-MSCs, allowing MSCs to migrate to the site of vascular injury, inhibit inflammation, reduce neointimal hyperplasia, and promote vascular repair.
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BACKGROUND: Current treatments for metabolic syndrome are comprehensive treatments with drugs, aiming to improve patient’s life. Patients are required to have a high compliance to follow-up and have developed various adverse reactions. Therefore, there is no treatment to fundamentally delay the development of metabolic syndrome. OBJECTIVE: To establish a tree shrew model of metabolic syndrome and evaluation techniques, and to explore the therapeutic effect of umbilical cord mesenchymal stem cells in the tree shrew model, providing theoretical basis and reference method for clinical application of stem cell transplantation in metabolic syndromes. METHODS: Umbilical cord mesenchymal stem cells of tree shrews were obtained by adherent culture method, and met the biological characteristics of umbilical cord mesenchymal stem cells. The dark red fluorescent iodide DIR was used to label tree umbilical cord mesenchymal stem cells. Thirty-two tree shrews were fed high-sugar, high-cholesterol, high-salt diet and syrup diet, and injected streptozotocin to make metabolic syndrome models. Animal models were randomly divided into model control group (n=10) and cell treatment group (n=22). In the cell treatment group, the umbilical cord mesenchymal stem cells labeled in vitro were injected into the tail vein, and the model group was injected an equal volume of physiological saline in the meanwhile. Blood biochemical indicators, glucose tolerance, insulin resistance index and arterial blood pressure were measured after transplantation. RESULTS AND CONCLUSION: The tree shrew model of metabolic syndrome was successfully constructed, showing obvious insulin resistance, hyperglycemia, lipid metabolism disorder, and hypertension, which met the diagnostic criteria of metabolic syndrome. Umbilical cord mesenchymal stem cells transplantation could significantly reduce the blood glucose and blood lipid levels, improve insulin resistance and regulate insulin secretion in the tree shrew model of metabolic syndrome. Transplanted umbilical cord mesenchymal stem cells could be homing to the liver, kidney and pancreas of the tree shrew model of metabolic syndrome, and produce certain repairing effects.
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BACKGROUND: Human umbilical cord mesenchymal stem cells (hUC-MSCs) have been drawing a great attention due to their potential therapeutic effect in a variety of diseases, including immune-mediated diseases. Further characterization of the immunomodulatory properties and action pathways of hUC-MSCs is necessary to ensure their safety and effectiveness in clinical application. OBJECTIVE: To investigate the immunomodulatory properties of hUC-MSCs. METHODS: HUC-MSCs were directly co-cultured with CFSE-labeled peripheral blood mononuclear cells (PBMCs) at the ratio of 1:5, 1:10, and 1:20, or indirectly co-cultured with CFSE-labeled PBMCs at the ratio of 1:5 via the Transwell co-culture system. Phytohemagglutinin- stimulated PBMC proliferation and the percentages of Th1, Th17 and Treg subgroups in the CD4+ T cells were determined by flow cytometry. The levels of tumor necrosis factor α and interferon γ were determined by ELISA. RESULTS AND CONCLUSION: After direct co-culture, hUC-MSCs significantly inhibited the phytohemagglutinin-stimulated PBMCs proliferation in a dose-dependent manner, whereas the inhibitory effect disappeared in the Transwell co-culture system. A significant decrease of Th1, Th17 cells and an increase of Treg cells were detected in the PBMCs co-cultured with hUC-MSCs compared to the PBMCs cultured alone. Furthermore, hUC-MSCs co-culture significantly reduced tumor necrosis factor α and interferon γ levels in the PBMCs. These findings indicate that cell-to-cell contact is essential for hUC-MSCs to inhibit the proliferation, differentiation and inflammatory factor secretion of immune cells.
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BACKGROUND: TDP43 may be a negative regulator of MAPK signaling pathway under hypoxic-ischemic conditions. However, its effect on JNK and p38 MAPK signaling pathways in osteoarthritis remains unclear.OBJECTIVE: To investigate the expression of chondrocyte lesion-related gene RACK1 in wild type TDP43 involved in osteoarthritis, and to analyze its stress effect. METHODS: Human umbilical cord mesenchymal stem cells were transfected by TDP43 lentivirus, and the ability to differentiate into chondrocytes in vitro was analyzed. Umbilical cord mesenchymal stem cells transfected by TDP43 lentivirus, empty vector and without transfection were co-cultured with chondrocytes for 12 days. The chondrocyte proliferation was detected at 0, 3, 6, 9 and 12 days of co-culture. The chondrocyte apoptosis rate was detected by flow cytometry at 3 days of co-culture. The expression levels of TDP43, RACK1, p38, JNK, AP-1 and cl-xl in chondrocytes were detected by qRT-PCR at 3 days of co-culture. RESULTS AND CONCLUSION: (1) After TDP43 lentivirus transfection, human umbilical cord mesenchymal stem cells could differentiate into chondrocytes. (2) The morphology of chondrocytes co-cultured with TDP43 lentivirus transfected-umbilical cord mesenchymal stem cells showed significant change, and the cells became large with abundant branches. Chondrocytes co-cultured with empty vector transfected- or non-transfected umbilical cord mesenchymal stem cells were spindle-shaped in appearance and showed adherent growth with no morphological changes. (3) After co-cultured with TDP43 lentivirus transfected-umbilical cord mesenchymal stem cells, the apoptosis of chondrocytes was promoted, and the cell proliferation was inhibited (P < 0.05). (4) The expression levels of TDP43, RACK1, p38, JNK, AP-1 and Bcl-xl in the chondrocytes co-cultured with TDP43 lentivirus transfected-umbilical cord mesenchymal stem cells were significantly higher than those in the chondrocytes co-cultured with non-transfected- and empty vector-transfected-umbilical cord mesenchymal stem cells. (5) To conclude, high expression of TDP43 in chondrocytes can activate the expression of RACK1, and further regulate chondrocyte proliferation and apoptosis.
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OBJECTIVE@#To evaluate the capacity and efficiency of human umbilical cord mesenchymal stem cells (HUCMSCs) to differentiate into neuron- like cells after induction with B27- supplemented serum- free medium.@*METHODS@#HUCMSCs at passage 4 were cultured for 14 days with serum-containing medium (SCM) (group A), SCM supplemented with 20 ng/mL nerve growth factor (NGF) and 10 ng/mL basic fibroblast growth factor (bFGF) (group B), serum-free medium (SFM) (group C), or SFM supplemented with 20 ng/mL NGF and 10 ng/mL bFGF. The culture medium were changed every 3 days and the growth of the neurospheres was observed using an inverted microscope. The cell markers were analyzed with flow cytometry and the expressions of nestin, neuron- specific enolase (NSE), neurofilament heavy polypeptide (NEFH), and glial fibrillary acidic protein (GFAP) were quantified by quantitative real-time PCR (qRT-PCR) and Western blotting.@*RESULTS@#Before induction, HUCMSCs expressed abundant mesenchymal stem cell surface markers including CD29 (99.5%), CD44 (49.6%) and CD105 (77.7%). Neuron-like cells were observed in the cultures on days 7, 10, and 14, and the cell differentiation was the best in group D, followed by groups C, B and A. In all the 4 groups, the cellular expressions of nestin and GFAP gradually lowered while those of NEFH and NSE increased progressively. The expressions of GFAP, NEFH, nestin and NSE were significantly different between group A and the other 3 groups ( < 0.001 or 0.05).@*CONCLUSIONS@#B27-supplemented SFM effectively induces the differentiation of HUCMSCs into neuron- like cells, and the supplementation with cytokines (NGF and bFGF) strongly promotes the cell differentiation.
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BACKGROUND: Human umbilical cord mesenchymal stem cells play a vital role in the repair of the blood-brain barrier after traumatic brain injury. OBJECTIVE: To investigate the protective effect of human umbilical cord blood mesenchymal stem cell transplantation on the blood-brain barrier after traumatic brain injury in rats and its possible mechanism. METHODS: Sixty Sprague-Dawley rats were randomly divided into sham operation group, injury control group (model group), cell transplantation group and Sunitinib group, with 15 rats in each group. Traumatic brain injury model was established by improved hydraulic impact method in all the groups except for the sham operation group. Rats in the sham operation group and model group were injected with 1 mL of normal saline, and those in the cell transplantation group were injected with 1 mL of 2×109/L human umbilical cord blood mesenchymal stem cells. The injection was done via the tail vein at 0.5, 24, and 48 hours after modeling. In the Sunitinib inhibitor group, the rats were given oral PDGFR-β pathway inhibitor, Sunitinib (80 mg/kg), from 1 day before modeling until being executed. Three days after modeling, the water content in brain tissue was measured by dry-wet specific gravity method, the permeability of the blood-brain barrier was measured by Evans blue method, expression of GFAP and vWF was observed by immunofluorescence staining and the expression of blood-brain barrier related proteins and PDGFR-β pathway proteins was detected by western blot method. RESULTS AND CONCLUSION: Compared with the sham operation group, the brain water content of the model group increased significantly (P < 0.05), while that of the cell transplantation group was significantly lower than that of the model group (P < 0.05). The Evans blue content in the model group was significantly higher than that in the sham operation group (P < 0.05), while the Evans blue content in the cell transplantation group was significantly lower than that in the model group (P < 0.05). Compared with the sham operation group, the expression of vWF and GFAP increased significantly in the model group (P < 0.05), while compared with the model group, the expression was significantly reduced in the cell transplantation group (P < 0.05). Western blot showed that ZO-1, Oclaudin-5, and PDGFR-β protein expressions in the model group were significantly lower than those in the sham operation group (P < 0.05), while these expressions were significantly increased in the cell transplantation group as compared with the model group (P < 0.05). To conclude, intravenous injection of human umbilical cord mesenchymal stem cells through the tail ein can reduce the permeability of blood-brain barrier and play a neuroprotective role in rats with traumatic brain injury. Its possible mechanism is related to the promotion of PDGFR-β expression in the injured area.
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@#A glioma, especially a grade IV glioblastoma, is a malignant tumour with a poor prognosis despite growing medical advancements. Researchers have been looking for better and more effective treatments targeting the molecular pathways of gliomas due to glioblastomas’ ability to develop resistance to chemotherapies. Moreover, glioma stem cells (GSC) contribute to maintaining the glioma population, which benefits from its ability to self-renew and differentiate. Recent research has reported that through the introduction of umbilical cord mesenchymal stem cells (UCMSC) into glioma cells, the growth and development of the glioma cells can be downregulated. It has more currently been found out that UCMSC release extracellular vesicles (EVs) containing miRNA that are responsible for this phenomenon. Therefore, this review analyses literature to discuss all possible miRNAs contained within the UCMSC’s EVs and to elaborate on their molecular mechanisms in halting gliomas and GSC growth. This review will also include the challenges and limitations, to account for which more in vivo research is suggested. In conclusion, this review highlights how miRNAs contained within UCMSC’s EVs are able to downregulate multiple prominent pathways in the survival of gliomas.
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Umbilical cord mesenchymal stem cells(UC-MSC)are a kind of stem cell group with high self-renewal ability and multi-directional differentiation potential. In recent years,studies have shown that UC-MSC is far more advan- tageous than the stem cells from other sources in terms of acquisition,storage and transplantation. UC-MSC also possesses the characteristics of secreting specific cytokines,playing an immunomodulatory role,or delivering”drugs”to the special tumor sites. Therefore,UC-MSC has become a promising tumor-targeting tool. This article reviews recent advances in the research on the antitumor mechanism and safety of UC-MSC.
ABSTRACT
Objective@#To explore the effect of the umbilical cord mesenchymal stem cells(UC-MSCs) on the pulmonary fibrosis in silicosis rats.@*Methods@#SPF male Sprague Dawley rats were randomly divided into control group, silica model group and UC-MSCs treatment group with 12 rats each group. SiO2 intra-tracheal injection(0.5 ml of 50 mg/ml/rat) were applied to silica model group and UC-MSCs treatment groups. After that UC-MSCs treatment group received 1 ml UC-MSCs suspension (3×106 cells/ml) by tail vein injection on the 29th, 36th, 43th and 50th day after exposure to the first silica suspension. On the 60th and 75th day after exposure to silica suspension, all animals were examed for pulmonary CT. Then the rats were euthanized on 75th day after the first exposure to silica.Lung's histopathological examination of the rats from all the groups were carried out. The content of hydroxyproline in lungs, TGF-β1 and IL-6 in serum were examined.@*Results@#The lung's histopathological examination showed no obvious inflammatory cell and no fibrosis in the lung tissue of the control group, there were a lot of inflammatory cell aggregation and collagen fiber deposition in silica model group, while in the UC-MSCs intervention group and treatment group, there were less inflammatory cells and collagen fiber. The rats from silica model groups had higher HYP, TGF-β1 and IL-6 than the rats from UC-MSCs treatment group and control group. Lung fields of rats in the control group were clear and no obvious high-density shadow. Different-sized granular high-density shadows or reticular fibrous shadows were found diffusely distributed in the lungs of the rats in silica model group. Lung field of rats in UC-MSCs intervention group and treatment group were less high density shadows, and more clear.@*Conclusion@#UC-MSCs can alleviate the pulmonary fibrosis in silica model rats through regulating the secretion of some fibrosis related cytokines.