ABSTRACT
Objective To investigate the effects of bone marrow mesenchymal stem cells (BMSC) on the expression of inflammatory factors in rats with multiple organ dysfunction syndrome (MODS). Methods BMSC extracted from the 4-week-old Sprague-Dawley (SD) rats was cultivated and identified in vitro, then the 4th passage of which was used in the experimental study. Sixty SD rats were randomly(random number) divided into three groups (n=20 in each group): Sham group (SG), MODS group (MG) and BMSC group (BG). Rats in the MG was injected by 1 mg/kg lipopolysaccaride (LPS) via great saphenous vein, rats in the SG injected with the same volume sterile phosphate buffer saline and rats in the BG infused by 1×106/cells BMSCs through the tail vein at 2 h after LPS injection. The survival rate, tissue pathological changes of the lung, liver and heart by hematoxylin and eosin (HE) staining, organ dysfunction measurement by blood gas analysis and biochemical indicators as well as the related inflammatory factors by protein microarray and enzyme linked immunosorbent assay (ELISA), were detected 72 h post operation. Multi-group quantitative data was analyzed by one way ANOVA, paired-comparisons by LSD-t test and the comparisons of survival curves in the three groups by Log-rank test. The value of P<0.05 was considered statistically significant. Results The survival rate in SG, MG and BG was 100％, 60％ and 80％, respectively. The survival curves showed that the survival rate of SG was higher than the MG and BG (SG vs. MG, χ2=9.798, P=0.0017; SG vs. BG, χ2=4.333, P=0.0374), but there was no significant difference comparing the BG to the MG (χ2=2.408, P=0.1207). The tissue congestion and edema, and inflammatory cells infiltration in the lung, liver, and heart of the MG were observed by HE staining, while these changes reduced in the BG. Compared with the SG, the levels of pH and PaCO2 and lactic acid (Lac) increased significantly (all P<0.01), the level of total bilirubin (TB) significantly increased [(0.801±0.501)U/L vs. (2.533±0.382)U/L, P=0.003], while the albumin(ALB) level decreased significantly[(35.471±4.015)U/L vs. (23.202±4.872)U/L, P<0.01], and creatine kinase (CK) level increased significantly in MG [(315.670±41.402) vs. (708.250±219.201), P=0.042]. After BMSC treatment, the organ function improved significantly (all P<0.05). Gamma interferon (IFN-γ) and monocyte chemotactic protein 1 (MCP-1) were the differential expression factors in protein chips. The results of ELISA were similar to the protein chips: compared with the SG, IFN-γ and MCP-1 expressions in the MG increased significantly (P<0.01). Compared with MG, the expressions of IFN-γ and MCP-1 decreased significantly in the BG (P<0.01). Conclusion BMSC administration could modulate the inflammatory response of MODS rats by inhibiting the levels of IFN-γ and MCP-1, and improve the organ function and the survival rate.
ABSTRACT
BACKGROUND:Co-transplantation of bone marrow mesenchymal stem cells (BMSCs) and hematopoietic stem cells (HSCs) can improve the survival rate following radiation damage. OBJECTIVE:To review the mechanisms of BMSCs in hematopoietic reconstitution and immunomodulation after radiation damage. METHODS: Relevant articles included in the Web of science from 2005 to 2016 were retrieved, and the keywords were mesenchymal stem cells; radiation; radiation injury; hematopoietic stem cells; hematopoietic reconstitution; immunomodulatory in English. Then we sorted and analyzed the articles which were closely related to the theme. RESULTS AND CONCLUSION:BMSCs can repair broken DNA double bonds by autophagy and HR/NHEJ pathway; support and protect HSCs by intercellular interactions and paracrine action; and repair radiation-induced hematopoietic injury by inhibiting inflammation and immune responses to maintain HSC homeostasis. Therefore, BMSCs may function as an effective treatment for radiation-induced hematopoietic injury.
ABSTRACT
BACKGROUND:Stem cell transplantation can, in theory, rebuild the immune system and regulate immunologic function. In addition, the transplanted stem cells may differentiate into tissues or cells we want. Therefore, this intervention may restore gastrointestinal inflammatory lesion and exert therapeutic effect on inflammatory bowel diseases (IBD). OBJECTIVE:To review the application of different kinds of stem cells in IBD treatment. METHODS:A computer-based search of CNKI, PubMed and FMJS was performed for articles concerning stem cel transplantation and IBD published after January 2002. The key words were stem cells, inflammatory bowel diseases, Crohn's diseases, ulcerative colitis, cell therapy in Chinese and English respectively. At last, 49 articles were included in result analysis. RESULTS AND CONCLUSION: Embryonic stem cells and adult stem cells both have certain therapeutic benefits in IBD. But which kind of stem cell is the safest and most efficacious cannot be confirmed, and at the same time the most effective and the optimal dose and way of transplantation of such cells are still unclear. All in all, there is still a lot of work to do to remove the obstacles.
ABSTRACT
Objective To investigate the regeneration and differentiation of HOCs in the 2-AAF/PHx rat models. To explore the expression of Cyr61and its mechanism in differentiation of HOCs in vitro. Methods In 2-AAF/PHx rats model,induction and expansion of HOCs were detected by immunochemistry and HE staining. West-ern blot was used for observing the expression of Cyr61. Furthermore,the expression of Cyr61 andβ-catenin were detected by Western blot in differentiation of WB-F344 cells in vitro. Results Cyr61 protein level increased as a re-sult of HOCs in rats livers after 2-AAF/PHx. In addition,the expression of Cyr61 and β-catenin significantly in-creased during WB-F344 cells differentiation in vitro. Conclusions Cyr61 might play an important role as a signal-ing mediator in HOCs response and closely correlate with Cyr61 andβ-catenin in proliferation and differentiation of HOCs.
ABSTRACT
BACKGROUND: Studies have shown that umbilical cord mesenchymal stem cells are ideal seed cells for tissueengineering research.OBJECTIVE: To isolate, culture and identify tree shrew umbilical cord mesenchymal stem cells, in order toestablish a standardized tree shrew umbilical cord mesenchymal stem cell lines.METHODS: Caesarean-isolated tree shrew umbilical cord samples were used to isolate and culture umbilical cordmesenchymal stem cells using tissue explant adherent method. Flow cytometry assay was used to detect cellsurface markers. Osteogenic and adipogenic induction media were used to induce umbilical cord mesenchymalstem cells to differentiate into osteoblasts and adipocytes.RESULTS AND CONCLUSION: The cultured umbilical cord mesenchymal stem cells expressed CD90 and CD105 with the positive rate of 99.9% and 99.8% respectively. Hematopoietic stem cell marker CD34 expression ratewas 0.0% and the endothelial cell marker CD31 expression rate was 0.7%, in line with the characteristics of umbilicalcord mesenchymal stem cell surface markers. Calcium nodules by alizarin red staining and lipid droplets by oil red Ostaining were observed in the induced cells. These experimental findings indicated that umbilical cord mesenchymalstem cells from tree shrews capable of osteogenic and adipogenic differentiation were successfully isolated and cultured.
ABSTRACT
Objective To investigate the underlying mechanism of bone marrow mesenchymal stem cells (BMSC) modulating the inflammatory response during the multiple organ dysfunction syndrome (MODS), especially the expression of inflammatory cytokines, which will provide new theoretical and experimental basis of MODS in clinic. Methods BMSC of Sprague-Dawley (SD) rat (female, 4 weeks) was extracted and cultivated, and the 4th passage were used in experimental study. According to the random number table, 60 female SD rats were divided into three groups (n = 20 per group): sham group, MODS group, BMSC group. MODS model in rats was induced by lipopolysaccaride (LPS, 1 mg/kg) via femoral vein injection. Sham group was injected with the sterile phosphate buffer saline (PBS) in the same volume. BMSC group, in which BMSC infusion was started at 2 hours after 0.5 mL LPS stimulation (1×106/cells) through the tail vein. The survival rate was observed after 72 hours in each group. Abdominal aortic blood was collected for routine blood and biochemical examination at 72 hours after operation. Protein microarray was used to detect the related 34 inflammatory cytokines. Signal ratio was defined as the differentially expressed factors when it was more than 2.0 or less than 0.5. And enzyme linked immunosorbent assay (ELISA) was be applied to validate the significant inflammation factor. Meanwhile, the heart, kidney, intestine tissue was harvested, then their pathological changes were observed by hematoxylin eosin (HE) staining.Results 20, 12, 16 rats lived in sham group, MODS group and BMSC group respectively at 72 hours after operation. Compared with the sham group, the indicators (routine blood, liver and kidney function, myocardial enzyme) were apparently unusual, and the heart, kidney, intestine tissue were injured obviously in the MODS group. After BMSC administration, the organ function was improved and tissue damaged was alleviated significantly. Protein microarray showed that interleukin-4 (IL-4) and receptor for advanced glycation end products (RAGE) were significantly different in 34 goal cytokines. The signal ratio change of IL-4 was 0.397, 1.124, 2.826 respectively, and the signal ratio of RAGE was 6.197, 1.552, 0.250, respectively in MODS/sham group, BMSC/sham group, BMSC/MODS group. ELISA validated the result that the expression level of IL-4 decreased significantly (ng/L:3.59±1.21 vs. 29.10±5.78) and the expression level of RAGE increased significantly (ng/L: 1.09±0.04 vs. 0.11±0.03) in MODS group as compared with sham group (bothP < 0.05). Compared with the MODS group, the level of IL-4 was obviously higher than that in BMSC group (ng/L: 9.59±2.21 vs. 3.59±1.21,P < 0.01), and RAGE decreased significantly (ng/L: 0.29±0.07 vs. 1.09±0.04,P < 0.05).Conclusions BMSC administration can regulate the expression of IL-4 and RAGE in the rats subjected to MODS. Moreover, BMSC can promote the restoration of tissue and organ function, thus improve the survival rate. BMSC may be the target in cell therapy for the inflammatory disease.
ABSTRACT
BACKGROUND: Mesenchymal stem cells attract extensive attention because of good biological characteristics and broad prospects, but the cells gradually show the characteristics of the aging with the increase of individual age or incubation time in vitro. Nonhuman primates have similar biological characteristics with human being, and have unique advantage in the animal model and disease treatment research.OBJECTIVE: To analyze the difference in proliferation and differentiation of bone marrow mesenchymal stem cells from macaques at different ages and to explore the effect of age on bone marrow mesenchymal stem cells and the possible mechanism.METHODS: Bone marrow samples from male macaques aged < 3 years and over 20 years were collected through bone marrow puncture, and divided into young group and elder group, with three macaques in each group. Then, bone marrow mesenchymal stem cells were isolated and cultured in vitro, and the morphological changes, proliferation and differentiation ability were observed. Age-related beta-galactosidase staining was performed, and protein microarray and ELISA were used to detect cytokine levels.RESULTS AND CONCLUSION: With age, the proliferation and differentiation of bone marrow mesenchymal stem cells from the elder macaques were reduced significantly, and the number of senescent cells increased significantly; the levels of interleukin-1b, interleukin-4, interleukin-6, tumor necrosis factor α and vascular endothelial growth factor were elevated obviously, the levels of heparin-binding basic fibroblast growth factor and placental growth factor were reduced. These findings indicate that the body's aging lead to the reduction in the proliferation, differentiation and cytokine secretion of bone marrow mesenchymal stem cells.
ABSTRACT
BACKGROUND:At present, there are few reports about the non-human primate models of type 2 diabetes mel itus in domestic and abroad, so it lacks of standardized production methods and evaluation criteria. OBJECTIVE:To establish a safe and effective type 2 diabetes mel itus model of rhesus monkey and evaluation method. METHODS:Twelve rhesus monkeys were randomly assigned to experimental group (n=9) and control group (n=3). Rhesus monkeys in the experimental group were fed with high-glucose and high-fat diet for 4 weeks, and intraperitoneal y injected with 30 mg/kg streptozotocin to establish models of type 2 diabetes mel itus. Rhesus monkeys in the control group were fed with an equal volume of physiological saline. At 12 weeks after injection, peripheral blood serum was col ected to measure fasting blood glucose, lipids, insulin, and C-peptide levels. Intravenous glucose tolerance test and C-peptide release test were used to detect pancreatic gland and pancreatic islet function. Histopathological examination was performed in pancreas, kidney and liver. RESULTS AND CONCLUSION:(1) 12 weeks after injection, fasting blood glucose, triglycerides, and total cholesterol levels were significantly higher in the experimental group than in the control group (P<0.05). Insulin and C-peptide levels were significantly lower in the experimental group than in the control group (P<0.05). (2) The area under the curve for intravenous glucose tolerance test was increased in the experimental group than in the control group (P<0.05). The area under the curve for C-peptide response test was significantly reduced in the experimental group than in the control group (P<0.05). (3) The pathological sections of pancreas, kidney and liver showed typical pathological changes of diabetes in the experimental group. (4) It is confirmed that we got high achievement about rhesus monkey models of type 2 diabetes mel itus made by high-glucose and high-fat diet combined with low-dose streptozotocin. It is a feasible, safe and effective method.
ABSTRACT
BACKGROUND:Tissue-engineered transplantation technique has become an ideal therapeutic regimen for degenerative disc diseases through reconstituting the biological functions of the degenerated intervertebral discs. OBJECTIVE:To construct a novel tissue-engineered annular fibrosus scaffold. METHODS:Konjac glucomannan, nano-hydroxyapatite and colagen were used to fabricate a new tissue-engineered annular fibrosus scaffold by wet spinning, chemical crosslinking, and freeze drying methods. Afterwards, X-ray diffraction and Fourier transform infrared spectrometer were used to analyze the scaffold qualitative components, physico-chemical property, biomechanical performance and cytocompatibility. RESULTS AND CONCLUSION:The bionic scaffold had a three-dimensional porous structure, with the average pore size of (425.8±47.3) μm, the average porosity of (73.4±5.6)%, and the water absorption of (718.6±24.3)%. In addition, the compressive strength of the scaffold was similar with that of the natural annular fibrosus. More importantly, the scaffold had good biocompatibility without cytotoxicity. These results show that the tissue-engineered annular fibrosus scaffold constructed by konjac glucomannan, nano-hydroxyapatite and colagen has proper three-dimensional porous structure, biocompatibility, porosity, water absorption and biomechanical strength.
ABSTRACT
BACKGROUND:If an extract can prolong the S phase and reduce the percentage of apoptosis after co-culture with cels, it is proved that the extract is able to promote cel proliferation. OBJECTIVE: To prove the effects of chicken egg-white extracts with different components on the proliferation, cel cycle and apoptosis of 293T cels. METHODS: An ultrafiltration unit was used to separate chicken egg-white extracts into different components that were > 10 ku, 3 ku and < 3 ku to co-culture with cels for 3 days. Then, cel proliferation, cel cycle and cel apoptosis were detected. RESULTS AND CONCLUSION:Chicken egg-white extract components of < 10 ku and < 3 ku could promote cel proliferation, increase the percentage of S-phase cels and reduce the percentage of apoptosis. In conclusion, chicken egg-white extract components that are < 10 ku and < 3 ku can promote cel proliferation, as wel as increase the percentage of S-phase cels and reduce apoptosis percentage.
ABSTRACT
Objective To test the effect of bone marrow mesenehymal stem cells (MSCs) transplantation on oxidative stress and the development of pulmonary emphysema in rats. Methods SD rats (n=26) were randomly divided into three groups:normal control group (group A, n=8),emphysema group (group B, n=8) and emphysema+MSCs transplantation group (group C, n=10).Rat models of emphysema was established by exposing rats to cigarette smoking for 14 weeks. Then rats of group C received MSCs transplantation. At the 14th and 28th days after 4 course of MSCs transplantations, one rat in group C was sacrificed at each time point and their lungs were preserved in frozen sections. Survival of MSCs in the lung tissues were observed by fluorescence microscopy. Eight weeks after transplantations, lung sections were stained by hematoxylin and eo?sin (HE) to observe the morphological alterations.Mean linear intercept (MLI) and mean alveolar numbers (MAN) were also measured. Serum and lung malondialdehyde (MDA) levels and superoxide dismutase (SOD) activity were also examined. Re?sults At the 14th day and 28th day after transplantations of MSCs, MSCs successfully localized to lung and survived in rat models of emphysema. Emphysematous changes of lung tissues were observed in both group B and group C. MLI was higher while MAN was lower in group B and C than those in group A (P<0.05). MLI and MDA levels in serum and lung were high?er while MAN level and SOD activity were lower in group B than those in group C (P<0.05).MDA levels in serum and lung was higher while SOD activity was lower in group B and C than those in group A (P<0.05). Conclusion MSCs transplanta?tions can effectively alleviates pulmonary emphysema in rat models which might through reducing oxidative stress .
ABSTRACT
Objective To establish a method for isolation of cynomolgus monkey umbilical cord mesenchymal stem cells.Methods Fresh cynomolgus monkey umbilical cord was directly minced into pasty fine pieces, and the pieces were cultured in tissue flask with DMEM/F12 medium supplemented with 10% fetal bovine serum.The morphological characteristics of the resulting cells were examined, and their expression of mesenchymal cell surface markers were analyzed by flow cytometry.The multidifferentiation potential was examined in vitro, too.Results The fibroblast-like cells were successfully isolated from the fresh umbilical cord by an adherent culture procedure.These adherent cells expressed mesenchymal markers including CD29, CD44, and CD90, and also could be induced to differentiate into adipocytes, osteoblasts and chondrocytes.Conclusion Mesenchymal stem cells can be isolated from fresh cynomolgus monkey umbilical cord by using an adherent culture procedure.
ABSTRACT
BACKGROUND:Systemic lupus erythematosus is an autoimmune disease characterized as an emergence of a variety of autoantibodies in serum and multi-system and multi-organ lesions. Currently, there is a lack of effective treatment options, and umbilical cord mesenchymal stem cel s are a promising therapy for systemic lupus erythematosus based on cel biological roles. OBJECTIVE:To observe the therapeutic efficacy of human umbilical cord mesenchymal stem cel transplantation in the treatment of systemic lupus erythematosus in mice. METHODS:Human umbilical cord mesenchymal stem cel s were isolated and cultured fol owed by labeling with DiR fluorescence. Experimental mice were divided into normal control group (C57BL mice), model control group (C57BL/lpr mice), low-, medium-and high-dose umbilical cord mesenchymal stem cel therapy groups (C57BL/lpr mice), with 10 mice in each group. Mice in the low-, medium-and high-dose groups were respectively injected 0.5×106, 1×106, 2×106 human umbilical cord mesenchymal stem cel s, once a week, for 3 consecutive weeks. At the end of treatment, blood samples were col ected to measure antinuclear antibody, anti-histone antibody, anti-double stranded DNA antibody changes;OPG and Foxp3 gene expression changes were detected by quantitative PCR method. RESULTS AND CONCLUSION:After treatment, the levels of anti-nuclear antibodies, anti-histone antibodies and anti-double stranded DNA antibodies in the peripheral blood of mice were al declined in the low-, medium-and high-dose groups, while the number of peripheral blood CD4+CD25+T cel s was significantly elevated. OPG and Foxp3 gene expression was also increased dramatical y in the low-, medium-and high-dose groups, which was similar to that in the normal control group and significantly different from that in the model control group (P<0.01). Experimental findings demonstrate that after transplantation of human umbilical cord mesenchymal stem cel s, al relevant indicators in C57BL/lpr mice recovered to the normal levels, and the high-dose treatment group had the most obvious effect.
ABSTRACT
AIM:To explore the effects of confluent endothelial progenitor cells (EPCs) derived from young and aged rats on the phenotype conversion, proliferation and migration of vascular smooth muscle cells ( SMCs) .METH-ODS:Mononuclear cells were obtained from the bone marrow of young (1~2 month old) and aged (19 to 26 month old) Sprague-Dawley rats and cultured with medium DMEM/F12 ( containing 15% fetal bovine serum, endothelial cell growth supplements (ECGs) 100 g/L, 1 ×105 units/L of penicillin and streptomycin, respectively).EPCs were characterized as double positive for DiI-Ac-LDL uptake and lectin binding.Abdominal aorta was obtained from 1 to 2 month old Sprague-Dawley rats.Vascular SMCs were cultured by tissue explant method and identified byα-SM-actin immunofluorescence.In transwell co-culture system, the confluent EPCs located in the upper chamber and SMCs were seeded on the lower cham-ber.The experiments were divided into passage 3 SMCs group (P3), passage 4 SMCs group (P4), passage 4 SMCs co-culture with EPCs derived from young rats group (P4YE) and passage 4 SMCs co-culture with EPCs derived from aged rats group (P4AE).The protein expression ofα-SM-actin and osteopontin was detected by Western blotting.[3H]-TdR incor-poration assay was used to determine the proliferation.SMC migration was analyzed by scratch wound healing assay.RE-SULTS:Compared with P3 group,α-SM-actin expression in P4 group significantly decreased and osteopontin protein ex-pression obviously increased, whereas no significant change was found in P4YE group.Compared with P4 group, confluent EPCs derived from young and aged rats both markedly increased α-SM-actin and decreased osteopontin expression in P4 SMCs.Compared with aged rat-derived EPCs, young rat-derived EPCs were more effectively to induce a delayed SMC phe-notype transition (from contractile phenotype to a synthetic phenotype), and to inhibit SMC proliferation and migration. CONCLUSION:Co-culture of confluent EPC induces a delayed vascular SMC phenotype transition and inhibits SMCs pro-liferation and migration.Young rat derived EPCs are more effective to induce a delayed vascular SMC phenotype transition and has stronger inhibitory effects on SMCs proliferation and migration compared with that derived from aged rats.
ABSTRACT
BACKGROUND:Tree shrew is a representative between insectivore and primates, has a high degree of evolution, is more inexpensive primates, has high use of medical biology, and has been attached by scholars. OBJECTIVE:To detect whether the commonly used secondary antibodies have immune response with tree shrew serum. METHODS:Western blot assay and enzyme linked immunosorbent assay were utilized to detect whether the tree shrew serum had cross-reacts with anti-rabbit, anti-goat, anti-human, anti-mouse, anti-rat, and anti-monkey secondary antibodies. RESULTS AND CONCLUSION:Western blot assay results indicated that tree shrew serums did not react with anti-rabbit, anti-goat, anti-human, anti-mouse, and anti-rat secondary antibodies and had cross reaction with anti-monkey secondary antibody. Enzyme linked immunosorbent assay results also indicated that tree shrew serums were cross-reactive with anti-monkey secondary antibody, but did not have cross-reactivity with the other secondary antibodies. Above data confirmed that the usual y soled secondary antibody cannot be used to immunoassay with tree shrews IgG. Only anti-monkey secondary antibody has cross-react with tree shrew serum. It is necessary to prepare anti-tree shrew IgG monoclonal and polyclonal antibodies. When no antibody is readily available at present, anti-monkey secondary antibody can be used to substitute detection, and can be widely applied in the study of tree shrew models of disease.
ABSTRACT
BACKGROUND:Traditional cel transplantation tracer methods require histological analysis and identification in vitro, which limits the clinical application of stem cel transplantation. So it is urgent to establish an in vivo noninvasive and repeatable tracer method. OBJECTIVE:To observe the effect of SPIO and DAPI double labeling on survival and proliferation of bone marrow mesenchymal stem cel s from macaques. METHODS:Bone marrow mesenchymal stem cel s were derived from bone marrow aspirates of healthy macaques using whole bone marrow adherence method. Then, the cel s were identified using flow cytometry detection. Bone marrow mesenchymal stem cel s were labeled using SPIO and DAPI. Fluorescent microscope was used to detect DAPI positive rate, and Prussian blue staining and transmission electron microscope were employed to measure SPIO positive rate. MTT assay was used to detect cel viability and proliferation. RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cel s were successful y isolated from healthy macaques using the whole bone marrow adherence method, and the cel purity was up to 95.1%. SPIO and DAPI were both successful to label the bone marrow mesenchymal stem cel s with a positive rate of 95%-98%, but had no influence on cel viability and proliferation.
ABSTRACT
Objective:To analyze the chicken egg-white extracts were co-cultured with cells whether elevated stem cells protein,whether the cells transformation into stem cell.Methods:Four kinds of cells,making a common culture,a 50% chicken egg-white extract co-cultured for 3 days,cells were collected and frozen at -80 degrees,sending the company to do stem cell protein microarray.Results:C57-BMSC has three proteins occurred statistically significant change , TS-UC-MSC has one proteins occurred a statistically significant change ,293T has one protein occurred a statistically significant change ,and 293T-GFP has one protein occurred a statistically significant change.Conclusion:50% chicken egg-white extract co-cultured cells,the cells occurred the phenomenon of transformation into stem cells.
ABSTRACT
BACKGROUND:Umbilical cord as childbirth waste has wide variety of sources and can be easily obtained, without any ethical and legal restrictions. Therefore, human umbilical cord mesenchymal stem cells break al the restrictions originated from other sources of mesenchymal stem cells. OBJECTIVE:To review the application of human umbilical cord mesenchymal stem cells in cartilage diseases, neuroglioma, ischemic brain injury, lung disease, liver disease and models of myocardial infarction. METHODS:The PubMed and Wanfang databases were searched by the first author using the keywords of“human umbilical cord mesenchymal stem cells, disease models, celltherapy”in English and Chinese, respectively. Seventy-three articles were searched and final y, 35 articles were included in result analysis. RESULTS AND CONCLUSION:Human umbilical cord mesenchymal stem cells have multilineage differentiation capacity similar to bone marrow mesenchymal stem cells. Compared with bone marrow mesenchymal stem cells, human umbilical cord mesenchymal stem cells have lower immunogenicity. Human umbilical cord mesenchymal stem cells show certain curative effects on cartilage disease, neuroglioma, ischemic brain injury, lung disease, liver disease and myocardial infarction, indicating that human umbilical cord mesenchymal stem cells can be used for celltransplantation to treat various diseases.
ABSTRACT
Objective To establish a method of identification of DKO mouse model of Duchenne muscular dystro-phy, and to assess the dystrophin regeneration after stem cell transplantation.Methods Heterozygous mice were mated and the resulting offspring were used to identify their genotype by SSP-PCR.The plasma creatine kinase level was measured by biochemical analyzer and histological changes in the DKO mice were analyzed using HE staining.Human umbilical cord mesenchymal stem cells were prepared and injected into the DKO mice hindlimb muscle, and dystrophin expression was de-tected by immunofluorescence staining at 2 months after injection.Results Mating of heterozygous mice generated three kinds of genotype offsprings, and 21.2%of the offsprings were identified as DKO genotype (285 bp) .DKO mice showed dystrophic symptoms, their plasma creatine kinase level was as high as 16988.52 ±617.48 IU/L, and significant histologi-cal changes including diverse myocyte sizes, numerous centrally nucleated cells and connective tissue proliferation or in-flammatory cells infiltration.Human dystrophin expression was detected in the DKO mouse hindlimb muscle at two months after injection of human umbilical cord mesenchymal stem cells.Conclusion DKO mouse genotype can be identified by SSP-PCR, and DKO mouse is an ideal animal model for studies of stem cell therapy for Duchenne muscular dystrophy.
ABSTRACT
BACKGROUND:Reprogramming somatic cells to generate pluripotent stem cells has a wide application in biomedical research. OBJECTIVE:To analyze the effect of different molecular weight proteins in chicken egg-white extract to elevate expression of pluripotent genes Oct-3/4 and Nanog in 293T cells. METHODS:The extracts of chicken egg-white were separated into more than 3 ku and less than 3 ku ingredients to be used for co-culture with 293T cells. There were four groups, 1×105 293T cells per wel , total 500μL. In the control group, 500μL culture medium was added;in the other three groups, 500μL chicken egg-white extract, more than 3 ku and less than 3 ku ingredients were respectively added. Quantitative PCR was used to determine the relative expression levels of pluripotent genes Nanog and Oct-3/4 in 293T cells. RESULTS AND CONCLUSION:By using co-culture method, more than 3 ku ingredients have a role to increase the expression of pluripotent genes Oct-3/4 and Nanog, but less than 3 ku ingredients cannot elevate the expression of pluripotent genes. This indicates that the ingredient of chicken egg-white extract to elevate the expression of pluripotent genes is more than 3 ku proteins.