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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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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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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
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: 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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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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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: 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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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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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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@#AIM: To investigate the effect of transplanted pigmented epithelium-derived factor(PEDF)on human umbilical cord-derived mesenchymal stem cells(hUCMSCs)in the treatment of retinitis pigmentosa in rat models. <p>METHODS: The hUCMSCs were isolated and cultured, hUCMSCs were transfected with PEDF recombinant lentivirus. Experimental royal college of surgeon(RCS)rats were randomly divided into 3 groups, 8 rats in each group. The experimental group was injected with PEDF-hUCMSCs subcutaneously. The hUCMSCs control group was injected with the same amount of hUCMSCs. The PBS control group was injected with the same amount of PBS. At 4 and 8wk after injection, electroretinography(ERG), the thickness of retinal outer nuclear layer(ONL)and green fluorescent protein(GFP)staining were observed. Immunofluorescence staining of retinal sections and the phagocytosis of MERTK protein were evaluated to determine phagocytosis. <p>RESULTS: PEDF gene carrying recombinant lentiviral vector could efficiently infect hUCMSCs. After infection, hUCMSCs were sub cultured and the lentivirus prolonged the expression in the cells of the target gene. At 8wk after transplantation, the amplitude of b-wave in ERG were significantly higher in the experimental group than in the control groups. At 4 and 8wk after transplantation, morphological changes of ONL thickness in the experimental group were significantly higher than those in the control groups(all <i>P</i><0.05). At 4 and 8wk after transplantation, hUCMSCs infected by lentiviral vector carrying PEDF were found survival in the subretinal cavity by GFP staining, and the PEDF-hUCMSCs were found having phagocytosis for outer segment fragments of photorecepter cells by immunofluorescence staining. At 4 and 8wk after transplantation, the Protein band gray values in experimental group were significantly higher than that in control groups, which indicated that the expressions of MERTK protein in experimental group were also increased significantly, and the phagocytic capacity of RPE were improved. <p>CONCLUSION: Transplantation of pigment epithelium-derived protein modified human umbilical cord-derived mesenchymal stem cells has a protective effect on the retina of RCS rats.
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[Objective]To investigate the effect of human umbilical cord mesenchymal stem cells on infected state of human alve?olar type Ⅱ epithelial cells.[Methods]Human alveolar type Ⅱ epithelial cells A549(1×105/mL)2 mL and PA(3×104 CFU/mL)2 mL has grown after 6 hours,add hUCMSC(1 × 106/mL)2 mL as the experimental group,add equal amounts of phosphate buffer (PBS)for infection,A549 and PBS and the medium has grown as the control group. A549 cells morphological changes between the compared groups(Transmission electron microscope,TEM),A549 cell viability(new CCK-8 cell proliferation assay Kit),A549 cells apoptosis(Annexin V-FITC/PI double staining flow cytometry)and the expression of A549 pulmonary surfactant A(SP-A) (Western blot).[Results]Transmission electron microscope cell morphology observation displayed ,infection group A549 cell dam?aged obviously,cell quality appeared empty bubble degeneration,chromatin height agglutination,visible apoptosis bodies;experi?ment group cell package film structure full,nuclear film full,nucleolus obviously,nuclear chromatin electronic density low,chroma? tin uniform,no apoptotic bodies;control group A549 cell structure full,membrane surface micro-fluff rich,nuclear film full,nucle?ar week clearance structure normal,chromatin uniform;infection group and control group compared,Infection group A549 cell sur?vival significantly reduced[(70.35±2.89)% and(97.37±2.07)%,n=3,P<0.01],apoptosis rate significantly increased[(8.63%± 0.16)%and(2.55±0.11)%,n=3,P<0.01],In the infected group,PA could damage A549 cells and decrease the amount of SP-A ex?pression(n=5,P<0.05). In the experiment group,the protective effect of hUCMSC on the A549 cells after infection may increase the expression of SP-A(n=5,P<0.05);[Conclusions]HUCMSC inhibits the infection of A549 cells apoptosis and protection of A549 cells secrete SP-A.
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Objective To find the seed cells for replacing bone marrow mesenchymal stem cells(BMSCs) to treat bone defects by compared the capacity of osteogenesis from the BMSCs, human umbilical cord mesenchymal stem cells(UC-MSCs) and human placental mesenchymal stem cells(P-MSCs). Methods Three MSCs were cultured in DMEM/Ham's F-12 medium containing 10% fetal bovine serum,cell proliferation curve was drawn by CCK8 detec-ted,and the cell surface antigens were measured using flow cytometry. Osteogenic ability was confirmed by the al-kaline phosphatase(ALP) staining,alizarin red staining. To further explore the difference in organic components, their underlying genotypes and proteins, including RUNX2, ALP and osteocalcin(OCN), were analyzed by RT-qPCR and Western blot. Results Cell growth curve analysis indicated that three MSCs were in the exponential stage at 3 days following incubation. Flow cytometry analysis showed that more than 98% cells were positive for CD44, CD90 and CD105. ALP and Alizarin red staining displayed that three MSCs presented good mineralizationg ability by osteogenesis induced medium. RT-qPCR and Western blot showed that the three MSCs experiment group higher expression levels of osteogenic markers including RUNX2, ALP and OCN during the initial 9 and 18 days when compared with control(P<0.05). Conclusions UC-MSCs and P-MSCs have good osteogenic ability, which may function as a potential bone tissue engineering seed cells for the treatment of bone defects.
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Objective To find the seed cells for replacing bone marrow mesenchymal stem cells(BMSCs) to treat bone defects by compared the capacity of osteogenesis from the BMSCs, human umbilical cord mesenchymal stem cells(UC-MSCs) and human placental mesenchymal stem cells(P-MSCs). Methods Three MSCs were cultured in DMEM/Ham's F-12 medium containing 10% fetal bovine serum,cell proliferation curve was drawn by CCK8 detec-ted,and the cell surface antigens were measured using flow cytometry. Osteogenic ability was confirmed by the al-kaline phosphatase(ALP) staining,alizarin red staining. To further explore the difference in organic components, their underlying genotypes and proteins, including RUNX2, ALP and osteocalcin(OCN), were analyzed by RT-qPCR and Western blot. Results Cell growth curve analysis indicated that three MSCs were in the exponential stage at 3 days following incubation. Flow cytometry analysis showed that more than 98% cells were positive for CD44, CD90 and CD105. ALP and Alizarin red staining displayed that three MSCs presented good mineralizationg ability by osteogenesis induced medium. RT-qPCR and Western blot showed that the three MSCs experiment group higher expression levels of osteogenic markers including RUNX2, ALP and OCN during the initial 9 and 18 days when compared with control(P<0.05). Conclusions UC-MSCs and P-MSCs have good osteogenic ability, which may function as a potential bone tissue engineering seed cells for the treatment of bone defects.
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Objective To investigate the effects of human umbilical cord mesenchymal stem cells (UC-MSCs ) on vascular endothelial growth factor ( VEGF ) and monocyte chemoattractant protein-1 ( MCP-1 ) of acute myocardial ischemia-reperfusion (AMI-R) injury in rats. Methods 24 Sprague-Dawley rats were randomly divided into sham group, AMI-R group and UCMSCs treatment groups on average. The rats were sacrificed on the 10th day after UCMSCs transplantation, and the myocardial tissues below the ligature were taken. The mRNA and protein expressions of MCP-1 of the tissue were detected by RT-PCR and Western Blot respectively, and the expression of VEGF protein was detected by immunohistochemistry. Results The relative expression levels of MCP-1 mRNA and the protein in UCMSCs group were significantly lower than those in sham group and AMI-R group (all P<0.05). The expression of VEGF protein in UCMSCs group was significantly higher than that in sham group and AMI-R group, the differences were statistically significant(all P<0.05). Conclusion UCMSCs transplantation can promote the angiogenesis and decrease the inflammation reaction in the treatment of acute myocardial ischemia-reperfusion injury.
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AIM: To compare the therapeutic effects of transplantation of human umbilical cord mesenchymal stem cells (hUCMSC) through different ways on diabetic mice.METHODS: hUCMSCs were labeled with enhanced green fluorescent protein (EGFP) and luciferase (Luc) reporter gene, and then the cells were transplanted into the diabetic mice through pancreas or tail vein to monitor the migration of the hUCMSCs in vivo.The pathological changes of pancreas tissue sections were determined by HE staining.Weight and blood glucose of the mice were measured dynamically.To compare the therapeutic effects, serum insulin levels were analyzed and glucose tolerance test were also performed.RESULTS: In vivo bioluminescence imaging results showed that the hUCMSCs transplanted into pancreatic capsule was mainly located in the pancreas while the hUCMSCs transplanted through vein tail injection was mainly located in the lung.HE staining illustrated that islet cells presented distinctive boundary and no infiltration of inflammatory cells in pancreatic capsule transplantation group was observed, but a little inflammatory cell infiltration and fibrosis formation in tail vein injection group were seen.A significant decrease in blood glucose level and a significant increase in serum insulin level in pancreas transplantation group were showed as compared with vein tail injection group.CONCLUSION: Transplantation of hUCMSCs through different approaches demonstrates different effects.The transplantation of hUCMSCs into pancreatic capsule is more effective on hyperglycemia reversion, insulin secretion and improvement of beta-cell function than that through tail vein.
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Recurrent laryngeal nerve (RLN) injury can result in unilateral or bilateral vocal cords paralysis, thereby causing a series of complications, such as hoarseness and dyspnea. However, the repair of RLN remains a great challenge in current medicine. This study aimed to develop human umbilical mesenchymal stem cells (HuMSCs) and nerve growth factor (NGF)-loaded heparinized collagen scaffolds (HuMSCs/NGF HC-scaffolds) and evaluate their potential in the repair of RLN injury. HuMSCs/NGF HC-scaffolds were prepared through incorporating HuMSCs and NGF into heparinized collagen scaffolds that were prefabricated by freeze-drying in a template. The resulting scaffolds were characterized by FTIR, SEM, porosity, degradation in vitro, NGF release in vitro and bioactivity. A rabbit RLN injury model was constructed to appraise the performance of HuMSCs/NGF HC-scaffolds for nerve injury repair. Electrophysiology, histomorphology and diagnostic proteins expression for treated nerves were checked after application of various scaffolds. The results showed that the composite scaffolds with HuMSCs and NGF were rather helpful for the repair of broken RLN. The RLN treated with HuMSCs/NGF HC-scaffolds for 8 weeks produced a relatively normal electromyogram, and the levels of calcium-binding protein S100, neurofilament and AchE pertinent to nerve were found to be close to the normal ones but higher than those resulted from other scaffolds. Taken together, HuMSCs/NGF HC-scaffolds exhibited a high score on the nerve injury repair and may be valuable for the remedy of RLN injury.
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Humans , Collagen , Dyspnea , Electrophysiology , Heparin , Hoarseness , In Vitro Techniques , Intermediate Filaments , Mesenchymal Stem Cells , Nerve Growth Factor , Paralysis , Porosity , Recurrent Laryngeal Nerve Injuries , Recurrent Laryngeal Nerve , Spectroscopy, Fourier Transform Infrared , Umbilical Cord , Vocal CordsABSTRACT
Objective: To study the antibacterial activity of human umbilical cord mesenchymal stem cells (hUCMSCs) culture supernatant and amoxicillin on Staphylococcus aureus isolated from burn wounds.
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Objective: To investigate the neuroprotective effects and mechanisms of Shennao Fuyuan Decoction (SFD) combined with implantation of human umbilical cord mesenchymal stem cells (hUC-MSCs). Methods: Rats were randomly divided into Sham operation group, model group[cerebral ischemia-reperfusion (I/R) model of rats prepared by middle cerebral artery occlusion, MCAO], SFD group (11.6 g/kg), hUC-MSCs group, combination group (SFD + hUC-MSCs), and these groups were separately divided into other three groups by execution time (days 4,8, and 15). The neurological function of rats was evaluated by neurological severity scores (NSS), the pathological changes of hippocampus were observed by HE staining, and the expression of BDNF and bFGF was detected by immunohistochemical and Werstern blotting method. Results: After cerebral ischemia, the neural necrocytosis of hippocampus in SFD group, hUC-MSCs group, and combination group was alleviated compared with model group; Compared with model group, the NSS was much lower and the expression of BDNF and bFGF was higher in SFD group (days 8 and 15), hUC-MSCs group, and combination group (days 4,8, and 15) (P<0.01 and 0.05); Compared with SFD group and hUC-MSCs group, the NSS was lower and the expression of BDNF and bFGF was higher in combination group (days 8 and 15) (P<0.05). Conclusion: The combination of SFD and hUC-MSCs implantation could significantly improve the restoration of nuerological defects after cerebral ischemia of rats, and this neuroprotective effect is probably due to the promotion of BDNF and bFGF expression.