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    05 February 2016, Volume 20 Issue 6 Previous Issue    Next Issue
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    Mechanism by which dihydroartemisinin inhibits invasion and migration of glioma stem cells
    Wu Yan-lin, Cai Zheng, Zhang Ming-zhi, Fu Xiao-rui
    2016, 20 (6):  765-770.  doi: 10.3969/j.issn.2095-4344.2016.06.001
    Abstract ( 266 )   PDF (508KB) ( 792 )   Save
    BACKGROUND: Dihydroartmisinin can promote apoptosis of glioma cells GL261, but its effect on glioma stem cells is still unknown.
    OBJECTIVE: To investigate the preliminary mechanism that dihydroartemisinin inhibits migration and invasion of glioma stem cells.
    METHODS: Glioma stem cells were isolated from mouse malignant glioma cell lines GL261. Immunofluorescence analysis was conducted to identify the characteristics of glioma stem cells. Migration and invasion abilities of glioma stem cells were analyzed by Transwell assay. The mRNA expressions of Toll-like receptor 2, matrix metalloproteinase-2 and matrix metalloproteinase-9 were examined by real-time fluorescence quantitative PCR.
    RESULTS AND CONCLUSION: The characteristics of glioma stem cells were identified by CD133 and Nestin staining. The migration and invasion of glioma stem cells were attenuated by dihydroartemisinin dose-dependently. Moreover, the mRNA expression of Toll-like receptor 2, matrix metalloproteinase-2 and matrix metalloproteinase-9 was also decreased by dihydroartemisinin in a dose dependent manner. These results suggest that dihydroartemisinin inhibits the migration and invasion of glioma stem cells probably through attenuation of Toll-like receptor signaling pathway. 
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    Effect of cholestatic serum on differentiation of bone marrow mesenchymal stem cells into hepatocytes
    Li Wei, Wang Lin, Zeng Jie, Chen Fang
    2016, 20 (6):  771-776.  doi: 10.3969/j.issn.2095-4344.2016.06.002
    Abstract ( 257 )   PDF (455KB) ( 319 )   Save
    BACKGROUND: Under certain conditions, bone marrow mesenchymal stem cells can be differentiated into hepatocytes, which are an important source of liver cells. Moreover, multiple factors can be involved in this induced differentiation process.
    OBJECTIVE: To investigate the inducible effect of cholestatic serum on the differentiation of bone marrow mesenchymal stem cells into hepatocytes.
    METHODS: Cholestatic animal model was prepared in rats to extract cholestatic serum. Bone marrow mesenchymal stem cells isolated from rats were divided into three groups and cultured in serum-free hepatocyte medium, serum-free hepatocyte medium plus cholestatic serum, serum-free hepatocyte medium plus normal serum, respectively.
    RESULTS AND CONCLUSION: The positive expression of alpha fetoprotein and keratin 18 and mass concentration of albumin were significantly higher in the serum-free hepatocyte medium plus cholestatic serum group than the other two groups (P < 0.05). These findings indicate that cholestatic serum has a certain inducible role in the differentiation of bone marrow mesenchymal stem cells into hepatocytes. 
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    Icariin protects against cyclophosphamide-induced obstacle of mouse bone marrow mesenchymal stem cells differentiating into osteoblasts
    Yang Zhi-lie, Wang Cheng-long, Zhao Dong-feng, Chang Jun-li, Yang Cheng, Yang Yan-ping, Wang Yong-jun
    2016, 20 (6):  777-784.  doi: 10.3969/j.issn.2095-4344.2016.06.003
    Abstract ( 325 )   PDF (558KB) ( 436 )   Save

    BACKGROUND: Osteoporosis caused by chemotherapy has become one of the serious side effects that impact the skeletal system. Icariin shows a strong anti-osteoporosis activity, which can have protective effect on osteoporosis induced by chemotherapy.
    OBJECTIVE: To study the protective effect and mechanism of icariin against cyclophosphamide-induced obstacle of mouse bone marrow mesenchymal stem cells differentiating into osteoblasts.
    METHODS: MTT assay and alkaline phosphatase (ALP) staining were used to determine the optimal protective concentration of icariin against cyclophosphamide-induced obstacle of mouse bone marrow mesenchymal stem cells differentiating into osteoblasts. mRNA expressions of osteoblast-specific transcription factors, OC, ALP, Runx2, and Wnt/β-catenin signaling pathway target genes, β-catenin, C-Myc, cyclin D1, were determined using RT-PCR method at different time after intervention with the optimal concentration of icariin. Expressions of Runx2, β-catenin, c-Myc, cyclin D1 regulated by the optimal concentration of icariin were detected using western blot assay at the protein level.
    RESULTS AND CONCLUSION: Cell viability and ALP activity decreased significantly in the cyclophosphamide group compared with the control group, but there was no significant difference in cell viability between icariin group and cyclophosphamide group. Icariin at 100 μmol/L showed the best protective effect against cyclophosphamide-induced obstacle of osteogenic differentiation of bone marrow mesenhymal stem cells. Compared with the control group, cyclophosphamide chemotherapy reduced the expressions of ALP, OC, Runx2 at mRNA level and Runx2 at protein level, weakened the expressions of β-catenin, cyclin D1 at mRNA level and active β-catenin, Cyclin D1, c-myc at protein level, and increased the expression of DKK1. Compared with the cyclophosphamide group, 100 μmol/L icariin increased the expression of osteoblast-specific transcription factors and Wnt/β-catenin signaling pathway genes at mRNA and protein levels, and reduced the expression of DKK1 protein. These results show that cyclophosphamide can lead to osteogenic differentiation disorder of mouse bone marrow mesenchymal stem cells, and in contrast, icariin shows a protective effect and its optimal intervention concentration is 100 μmol/L. Additionally, the protective role of icariin is probably related to activation of Wnt/β-catenin signal pathway.
     

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    Effects of over-expression of Notch1 intracellular domain on the differentiation of c-Kit+ bone marrow mesenchymal stem cells
    Ha Yan-ping, Wang Zhen-liang, Lei Hong, Ding Ran-ran, Jiang Xiao-fan, Wang Ke-ke, Shen Zhia-hua, Jie Wei
    2016, 20 (6):  785-792.  doi: 10.3969/j.issn.2095-4344.2016.06.004
    Abstract ( 373 )   PDF (717KB) ( 341 )   Save

    BACKGROUND: Activation of Notch signaling plays a critical role in stem cell differentiation, and this effect seems to be cell-type dependent. Little is reported on the role of activation of Notch1 signaling in the differentiation of c-Kit+ bone marrow mesenchymal stem cells.
    OBJECTIVE: To analyze the influence of activation of Notch1 signaling on the differentiation of c-Kit+ bone marrow mesenchymal stem cells.
    METHODS: The Notch1 intracellular domain (N1-ICD) was obtained from the cDNA library by PCR and cloned into BamHI/AgeI digested adenoviral GV314 plasmid to construct N1-ICD overexpressing shuttle plasmid, and the positive clones were verified by sequencing. N1-ICD shuttle plasmid and helper plasmids pBHGloxΔE1,3 Cre were used to co-transfect HEK293T cells to obtain N1-ICD overexpressing adenoviral particles (N1-ICD-Ad). The c-Kit+ subpopulation were isolated from bone marrow mesenchymal stem cells of the Sprague-Dawley rat femur via magnetic activated cell sorting. After transfection of the c-Kit+ BMSCs with N1-ICD-Ad adenovirus, we assessed the activation of Notch1 signaling and differentiation in c-Kit+ bone marrow mesenchymal stem cells by quantitative RT-PCR and immunofluorescent staining.
    RESULTS AND CONCLUSION: N1-ICD coding sequence was successfully generated from the cDNA library, and then was cloned into the linearized adenoviral vectors GV314. The resistant clones were verified by sequencing. With the assistance of packaging plasmids, recombinant N1-ICD-Ad adenovirus plasmids were successful packaged in HEK293T cells, and its title was 2×1012 PFU/L. c-Kit+ bone marrow mesenchymal stem cells with the purity of 91.6% were successfully isolated from the bone marrow mesenchymal stem cells of the Sprague-Dawley rat femur. Compared with the blank and negative controls, N1-ICD-Ad infection in the c-Kit+ bone marrow mesenchymal stem cells led to substantial accumulation of N1-ICD in the cytoplasm and nuclei, significantly unregulated expressions of Hes1 (a downstream gene of Notch) and cardiomyocyte differentiation genes Nkx2.5 and cTnT, significantly increased the expression of von Willebrand factor, an endothelial cell differentiation gene, and mildly increased the expression of smooth muscle 22α, a smooth muscle cell differentiation gene. These experimental results indicate that the activation of Notch1 signaling contributes to multi-lineages differentiation of c-Kit+ bone marrow mesenchymal stem cells, and the construction of N1-ICD overexpressing adenoviral vector makes the foundation for further research on the role of Notch1 signaling in stem cell biology. 

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    Migration and distribution of superparamagnetic iron oxide-labeled adipose-derived stem cells in the brain of cerebral infarction rats: MRI observation
    Chen Long-hua
    2016, 20 (6):  793-798.  doi: 10.3969/j.issn.2095-4344.2016.06.005
    Abstract ( 352 )   PDF (503KB) ( 429 )   Save
    BACKGROUND: To trace the survival status of transplanted stem cells and the integration with host tissues using non-invasive imaging techniques are the focus of research in recent years.
    OBJECTIVE: To observe the distribution and migration of superparamagnetic iron oxide (SPIO)-labeled adipose-derived stem cells in the brain of rats with cerebral infarction.
    METHODS: Rat models of cerebral infarction were established and randomized into SPIO-labeled group and unlabeled group. At 1 day after modeling, the rats in the two groups were given SPIO-labeled adipose-derived stem cell suspension (10 μL) and unlabeled adipose-derived stem cell suspension (10 μL) into the brain, respectively. At 1, 7, 14 days after cell transplantation, neurological severity scores were measured, and MRI was used to observe the distribution of SPIO-labeled adipose-derived stem cells.
    RESULTS AND CONCLUSION: At 7 and 14 days after transplantation, the neurological severity scores in the two groups were significantly lower than those at 1 day after transplantation (P < 0.05). However, there was no difference in the neurological severity scores between the two groups (P > 0.05). At 14 days after transplantation, MRI findings showed low signals in the transplanted region, indicating the cells migrated from the corpus callosum to the lesion. These findings suggest that intracerebral transplantation of adipose-derived stem cells can promote neurological recovery from cerebral infarction in rats, and MRI can be used to visualize the distribution and migration of SPIO-labeled adipose-derived stem cells in the brain. 
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    Transplantation of human umbilical cord mesenchymal stem cells derived at different gestational weeks improves heart function in myocardial infarction models
    Wang Wei, Li Xiao-fu, Li Zhong-jian
    2016, 20 (6):  799-806.  doi: 10.3969/j.issn.2095-4344.2016.06.006
    Abstract ( 230 )   PDF (594KB) ( 488 )   Save

    BACKGROUND: Stem cells have multi-directional differentiation and self-replication abilities, under certain conditions, which can differentiate into myocardial cells to repair the damaged myocardium.
    OBJECTIVE: To investigate the effects of transplantation of umbilical cord mesenchymal stem cells derived at different gestational weeks on infarct size and angiogenesis in the infarct region of experimental rabbits with myocardial infarction.
    METHODS: Ten full-term umbilical cord samples and 10 umbilical cord samples of aborted fetuses at 10-12 gestation weeks were selected to in vitro isolate umbilical cord mesenchymal stem cells that were subjected to BrdU labeling. HLA-G expression was detected in the cells. Thirty white rabbits were selected to make myocardial infarction models, and 2 weeks after modeling, the model rabbits were randomized into aborted cell transplantation group, full-term cell transplantation group and control group (n=10 per group). Then, BrdU-labeled cells were injected correspondingly into the infarct region of rabbits in the two cell transplantation groups. Rabbits in the control group were subjected to an equal volume of serum-free. Four weeks after transplantation, heart function of rabbits was monitored using electrocardiogram, and myocardial tissues were taken to measure infarct size and blood capillary density.
    RESULTS AND CONCLUSION: HLA-G expression was different in different sources of umbilical cord mesenchymal stem cells: high HLA-G expression was found in the aborted umbilical cord mesenchymal stem cells, and meanwhile, low HLA-G expression was found in the full-term umbilical cord mesenchymal stem cells. Compared with the control group, the left ventricular end-diastolic volume and left ventricular ejection fraction of aborted and full-term cell transplantation groups were significantly improved, especially in the aborted cell transplantation group (P < 0.05). BrdU-positive cells were found in the infarct site in both transplantation groups. Compared with the control group, the infarct size and capillary density were improved most significantly in the aborted cell transplantation group followed by the full-term cell transplantation group (P < 0.05). Electrocardiogram findings showed significant improvement in both cell transplantation groups compared with the control group (P < 0.05), especially in the aborted cell transplantation group. These findings indicate that umbilical cord mesenchymal stem cells derived at low gestational weeks improve the heart function more significantly than the full-term umbilical cord mesenchymal stem cells, which have the potential to become a better source of cardiomyocytes for transplantation.

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    Insulin promotes the osteogenic differentiation of umbilical cord mesenchymal stem cells
    Zheng Song-hao, Ha Cheng-zhi, Yang Xu, Wang Yuan-he, Tian Shao-qi, Sun Kang
    2016, 20 (6):  807-813.  doi: 10.3969/j.issn.2095-4344.2016.06.007
    Abstract ( 252 )   PDF (395KB) ( 591 )   Save

    BACKGROUND: How to effectively and rapidly induce the osteogenic differentiation of human umbilical cord mesenchymal stem cells is the focus of the current stem cell research. Increasing evidence has demonstrated some growth factors, such as bone morphogenetic protein-2, have important effects on the transdifferentiation of umbilical cord mesenchymal stem cells into osteoblasts in vitro. However, widespread use of growth factors is limited because of high cost. Insulin is widely used in the cell culture and induction, but there is no report about the effect of insulin on the osteogenic differentiation of human umbilical cord mesenchymal stem cells.
    OBJECTIVE: To observe the effect of insulin on osteogenic differentiation of human umbilical cord mesenchymal stem cells and to explore the feasibility of human umbilical cord mesenchymal stem cell transplantation in the treatment of diabetic delayed fracture healing.
    METHODS: The passage 3 human umbilical cord mesenchymal stem cells were inoculated in two flasks, denoted as experimental group and control group. The insulin (10-7 mmol/L) was added to the experimental group but not to the control group. The proliferative capacity of human umbilical cord mesenchymal stem cells was evaluated by cell count kit-8 and alkaline phosphatase activity. The osteogenic differentiation capacity of human umbilical cord mesenchymal stem cells was evaluated by measuring the protein and mRNA expressions of type I collagen as well as osteocalcin mRNA level.
    RESULTS AND CONCLUSION: After 1-2 weeks of induction, compared with the control group, insulin could significantly increase the number of human umbilical cord mesenchymal stem cells in the experimental group, the activity of alkaline phosphatase and expressions of type I collagen osteocalcin mRNA (P < 0.05). These data indicate that insulin can promote the proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells.
     

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    Allogeneic bone marrow mesenchymal stem cell transplantation can improve the function of the aging heart
    Li Yan-ju, Ding Yuan-ting, Zhou Yuan, Wang Fei-qing, Zeng Qiang-wu, An Shi-gang, Liu Yang
    2016, 20 (6):  814-819.  doi: 10.3969/j.issn.2095-4344.2016.06.008
    Abstract ( 353 )   PDF (517KB) ( 307 )   Save

    BACKGROUND: Bone marrow mesenchymal stem cells can secrete a variety of factors in the local lesion, and these factors can promote cell proliferation and inhibit cell apoptosis.
    OBJECTIVE: To observe the curative effect of bone marrow mesenchymal stem cell transplantation on the aging heart of rats and to explore the possible mechanism of action.
    METHODS: Thirty Sprague-Dawley rats were randomized into three groups: normal blank group, model group and treatment group. Aging models were made in the latter two groups by injection of D-galactose. Rats in the treatment group were given allogeneic bone marrow mesenchymal stem cell injection, once a week, totally four times. At 1 week after final injection, the heart tissues were sliced into sections to observe the pathological changes using hematoxylin-eosin staining. Western blot assay was used to detect the expression of basic fibroblast growth factor in the heart tissues. Real-time PCR was used to measure the expression of p53 mRNA in the heart tissues.
    RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cell transplantation could improve the pathological morphology of the aging heart. Compared with the model group, the expression of basic fibroblast growth factor in the heart tissues was significantly higher in the treatment group (P < 0.05), but the mRNA expression of p53 was lower (P < 0.05). It is speculated that bone marrow mesenchymal stem cells can interact with heart cells to secrete basic fibroblast growth factor and reduce p53 mRNA expression, thereby playing a curative effect on the aging heart.
     中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

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    Transplantation of human telomerase reverse transcriptase-transfected human amniotic mesenchymal stem cells in the treatment of pulmonary hypertension
    Zhang Chao, Cao Jie, Zhang Su-fang
    2016, 20 (6):  820-826.  doi: 10.3969/j.issn.2095-4344.2016.06.009
    Abstract ( 288 )   PDF (534KB) ( 344 )   Save

    BACKGROUND: The development of stem cell transplantation and genetic modification technology provides new ideas and methods for the treatment of pulmonary hypertension.
    OBJECTIVE: To investigate the therapeutic effects of transplantation of human telomerase reverse transcriptase (hTERT)-transfected human amniotic mesenchymal stem cells in pulmonary hypertension rats.
    METHODS: Human amniotic mesenchymal stem cells were cultured and purified in vitro, and then transfected with adenovirus-medicated hTERT. Sixty-six adult Wistar rats were enrolled to prepare pulmonary hypertension models through intraperitoneal injection of 60 mg/kg monocrotaline and then 63 model rats were randomly assigned into three groups: model group treated with transplantation of 1 mL of L-DMEM via the jugular vein, cell transplantation group treated with transplantation of 1 mL of untransfected human amniotic mesenchymal stem cell suspension, and transfection group treated with transplantation of 1 mL of human amniotic mesenchymal stem cell suspension transfected with hTERT. Hemodynamic changes, plasma endothelin-1 level, hypertrophy index of the right ventricle and myocardial cell apoptosis were compared among different groups at 3 weeks after transplantation.
    RESULTS AND CONCLUSION: After 3 weeks of treatment, there were no differences in the arterial blood pressure of the three groups (P > 0.05); however, the systolic pressure of the pulmonary artery and mean pulmonary arterial pressure were significantly lower in the transfection group than the model group and cell transplantation group (P < 0.05). Hypertrophy index of the right ventricle had no difference among the three groups (P > 0.05). The level of plasma endothelin-1 was significantly lower in the transfection group than the model group and cell transplantation group (P < 0.05). Apoptosis in myocardial cells was significantly reduced in the transfection group compared with the model group and cell transplantation group (P < 0.05). Experimental findings suggest that the transplantation of hTERT-transfected human amniotic mesenchymal stem cells can improve the hemodynamic levels in pulmonary blood vessels of pulmonary hypertension rats to protect vascular endothelial cells and reduce myocardial cell apoptosis. 

     

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    Adipose-derived mesenchymal stem cell transplantation protects the myocardial tissues from acute myocardial infarction
    Wang Li-zhong, Li Yan, Mu Huai-bin, Huang Kun, Gao Jun
    2016, 20 (6):  827-833.  doi: 10.3969/j.issn.2095-4344.2016.06.010
    Abstract ( 239 )   PDF (519KB) ( 426 )   Save

    BACKGROUND: Adipose-derived mesenchymal stem cells have rich sources that are easily obtained, which can be used to treat acute myocardial infarction.
    OBJECTIVE: To investigate the therapeutic effect of adipose-derived mesenchymal stem cell transplantation on acute myocardial infarction in rats.
    METHODS: Rat models of acute myocardial infarction were made and subjected to adipose-derived mesenchymal stem cell transplantation in comparison with model and control (sham operation) groups.
    RESULTS AND CONCLUSION: Echocardiography findings showed significant improvement in the left ventricular end-systolic diameter, left ventricular end-diastolic diameter and ejection fraction in the cell transplantation group compared with the model group (P < 0.05). Hematoxylin-eosin staining showed that myocardial infarction was evident in the model group, in which, there were rarely viable myocardial tissues and few vessels in the infarcted region, but in the cell transplantation group, there were evident survived myocardial tissues and transplanted cells. The percentage of infarct size was significantly lower in the cell transplantation group than the model group (P < 0.05). Immunohistochemical staining showed that adipose-derived mesenchymal stem cells were able to survive in the infarcted myocardial tissues, and the expression of cardiac troponin T in the cell transplantation group was significantly higher than that in the model group (P < 0.05). Experimental data show that adipose-derived mesenchymal stem cell transplantation can protect the myocardial tissues after myocardial infarction, and effectively improve the myocardial function.  

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    Transplantation of Foxc2-transfected bone marrow mesenchymal stem cells for experimental femoral head necrosis in rabbits
    Ren Yi-de, Zhang Ya-feng, You Wu-lin, Wang Jian-wei
    2016, 20 (6):  834-840.  doi: 10.3969/j.issn.2095-4344.2016.06.011
    Abstract ( 254 )   PDF (577KB) ( 363 )   Save

    BACKGROUND: Core decompression can delay early osteonecrosis of the femoral head, but cannot completely repair the necrotic femoral head. Eventually, femoral head collapse, even bone necrosis, will occur.
    OBJECTIVE: To explore the curative effect of implantation of gelatin sponge carrying Foxc2-transfected bone marrow mesenchymal stem cells on the repair of experimental femoral head necrosis in rabbits.
    METHODS: Forty New Zealand white rabbits were selected, and femoral head necrosis models were prepared successfully in 24 of 40 rabbits. Then, model rabbits were randomized into four groups: blank control group (n=4) with no treatment, core decompression group (n=4), GFP group (n=8) subjected to core decompression and implantation of gelatin sponge carrying GFP-transfected bone marrow mesenchymal stem cells, and Foxc2 group (n=8) subjected to core decompression and implantation of gelatin sponge carrying Foxc2-transfected bone marrow mesenchymal stem cells. At 1, 2, 4 weeks after implantation, ELISA was used to detect Foxc2 protein levels in the transplanted region. At 4, 8, 12 weeks after implantation, MRI scan of the hip was performed, and femoral head tissues were taken and sliced into sections for hematoxylin-eosin staining to observe bone growth. At 12 weeks after implantation, histomorphometry measurement and transmission electron microscope observation were carried out.
    RESULTS AND CONCLUSION: At 1, 2, 4 weeks after implantation, Foxc2 was highly expressed in the femoral head in the Foxc2 group, which was significantly higher than that in the GFP group. At 4, 8, 12 weeks, only a few of new bone formed in the core decompression group and GFP group; at 12 weeks, fibrous tissues formed in the decompression channel. New bone formation was evident in the Foxc2 group, and at 12 weeks, the necrotic region was repaired completely. MRI findings showed normal femoral head morphology and signals in the Foxc2 group at 12 weeks, but there were decreased signals of the femoral head in the core decompression group and GFP group. These findings indicate that Foxc2-transfected bone marrow mesenchymal stem cell transplantation via core decompression has good curative effects on experimental femoral head necrosis in animals.
     

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    Bone marrow mesenchymal stem cell transplantation via the tail vein for liver fibrosis
    Wang Chang-wu, Fang Xin-hui
    2016, 20 (6):  841-847.  doi: 10.3969/j.issn.2095-4344.2016.06.012
    Abstract ( 263 )   PDF (614KB) ( 443 )   Save
    BACKGROUND: Liver fibrosis is the early stage of terminal liver diseases. Effective treatment for liver fibrosis can prevent the occurrence of terminal liver diseases. Bone marrow mesenchymal stem cell transplantation is a promising method to treat liver fibrosis.
    OBJECTIVE: To study the therapeutic effect of bone marrow mesenchymal stem cells on liver fibrosis in rats.
    METHODS: Eighteen Sprague-Dawely rats were randomized into three groups: control, model and cell transplantation groups. Animal models of carbon tetrachloride-induced liver fibrosis were made in the latter two groups. After modeling, 1 mL bone marrow mesenchymal stem cells (5×105) or the same volume of normal saline was injected via the tail vein into the rats in the cell transplantation and model groups, respectively. Rats in the control group were given no treatment. Degree of liver fibrosis, liver function, histological changes of the liver were detected and observed in the three groups at 4 weeks after treatment.
    RESULTS AND CONCLUSION: In the control group, the liver tissues had normal structure with no fibrosis; in the model group, proliferation of fibrous tissues in the portal area of the liver, inflammatory cell infiltration, vacuolar degeneration and irregular arrangement of liver cells, and tissue structure damage were observed; in the transplantation group, liver tissue damage was severer than the control group but milder than the model group. Levels of serum hyaluronidase, type IV collagen and procollagen III were significantly lower in the cell transplantation group than the model group (P < 0.05). These findings indicate that bone marrow mesenchymal stem cell transplantation can alleviate liver fibrosis and improve liver function in rats with carbon tetrachloride-induced liver fibrosis. 
     
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    Adipose-derived mesenchymal stem cell transplantation via the hepatic artery for the treatment of advanced liver diseases
    Guo Xian-li, Liu Yue, Zhou Li-min, Hu Yue
    2016, 20 (6):  848-854.  doi: 10.3969/j.issn.2095-4344.2016.06.013
    Abstract ( 261 )   PDF (569KB) ( 436 )   Save

    BACKGROUND: Stem cell transplantation is a promising treatment for advanced liver diseases, and adipose-derived mesenchymal stem cells are a hot topic following bone marrow mesenchymal stem cells.
    OBJECTIVE: To explore the therapeutic effect of adipose-derived mesenchymal stem cells transplantation via the hepatic artery on advanced liver diseases in rats.
    METHODS: Forty-five rats were randomized into three groups, 15 rats in each group: control group, model group and transplantation group. Rat models of liver cirrhosis were made in the latter two groups through subcutaneous injection of carbon tetrachloride. Then, 1 mL of CFSE-labeled adipose-derived mesenchymal stem cells was infused via the hepatic artery in the transplantation group, and the same volume of normal saline was infused in the model group. Control group had no treatment. Pathological changes, liver function and degree of hepatic fibrosis were observed in the three groups at 4 weeks after treatment.
    RESULTS AND CONCLUSION: After transplantation, green fluorescence-labeled adipose-derived mesenchymal stem cells were seen in the liver of rats. Hematoxylin-eosin staining and Masson staining showed unclear hepatic lobule structure in the model group with the formation of false lobules, cell cloudy swelling and loose, some degeneration and necrosis, and inflammatory cell infiltration; in the control group, there was nothing abnormal in the liver tissues of rats in the control group; in the transplantation group, the pathological changes of the rat liver were better than those in the model group, but worse than those in the control group. Compared with the model group, the level of serum albumin was higher in the control and transplantation group (P < 0.05), and the levels of bilirubin, aminotransferase and type IV collagen were lower in the control and transplantation group (P < 0.05). Thus, it can be seen that adipose-derived mesenchymal stem cell transplantation can improve liver function and reduce liver fibrosis in cirrhotic rats. 

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    Transplantation of bone marrow mesenchymal stem cells to improve blood glucose and urinary total protein levels in diabetic nephropathy rats
    Du Jun-wen, Wu Tao, Zhang Kun, Su Bai-yu, Lu Cai-ping, Wang Wei-chao, Lei Lin, Guo Jing-xia
    2016, 20 (6):  855-860.  doi: 10.3969/j.issn.2095-4344.2016.06.014
    Abstract ( 290 )   PDF (462KB) ( 357 )   Save

    BACKGROUND: Common strategies for preventing diabetic nephropathy include effective control of blood sugar and blood pressure, inhibition of the rennin-angiotensin system and lipid-lowering therapy, but it is often difficult to get the desired results.
    OBJECTIVE: To investigate the effect of transplantation of bone marrow mesenchymal stem cells on levels of blood glucose and urinary total protein in diabetic nephropathy rats.
    METHODS: Forty-five Sprague-Dawley rats were randomly divided into three groups (n=15 per group): normal control group, diabetic nephropathy group and stem cell transplantation group. Rats in the diabetic nephropathy and stem cell transplantation groups were given single use of 60 mg/kg streptozotocin to make diabetic nephropathy models. The same dose of citric acid-sodium citrate buffer was injected in the normal control group. After modeling, 200 μL of bone marrow mesenchymal stem cell solution (2×106) was injected into the left ventricle of rats in the stem cell transplantation group, and then at 7 days after the first transplantation, the cell transplantation was conducted again. The same dose of serum-free L-DMEM was injected intracardially into the rats in the normal control and diabetic nephropathy groups. Levels of urinary total protein and blood glucose were detected. 
    RESULTS AND CONCLUSION: At 1, 4, 8 weeks after treatment, the urinary total protein and blood glucose levels were significantly higher in the stem cell transplantation group and diabetic nephropathy group than the normal control group (P < 0.05). At 1 week after treatment, the urinary total protein and blood glucose levels were significantly lower in the stem cell transplantation group than the diabetic nephropathy group (P < 0.05). At 4 and 8 weeks after treatment, the total urinary protein and blood glucose levels were slightly higher in the diabetic nephropathy group than the stem cell transplantation group, but there was no significant difference (P > 0.05). These findings indicate that bone marrow mesenchymal stem cell transplantation in diabetic nephropathy rats can get good results in a short period, significantly improve the blood glucose and urinary total protein levels, but the long-term treatment effect is poor. 

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    Bone marrow mesenchymal stem cell transplantation protects against intestinal ischemia-reperfusion injury in rats
    Liu Hong-feng, Li Lu
    2016, 20 (6):  861-867.  doi: 10.3969/j.issn.2095-4344.2016.06.015
    Abstract ( 212 )   PDF (632KB) ( 339 )   Save

    BACKGROUND: Bone marrow mesenchymal stem cells have good proliferation and paracrine functions, which have irreplaceable advantages in the treatment of intestinal diseases.
    OBJECTIVE: To explore the effects of bone marrow mesenchymal stem cell transplantation on intestinal ischemia-reperfusion injury in rats.
    METHODS: Forty-eight Sprague-Dawley rats were enrolled to make animal models of ischemic reperfusion injury of the intestine, and then model rats were randomized into experimental and control groups. After modeling, 1 mL bone marrow mesenchymal stem cells or the same volume of normal saline were injected into the intestinal mucosa of rats in the two groups, respectively. At hours 0, 2, 6, 24, 72, 120 after injection, serum diamine oxidase, tumor necrosis factor α, and D-lactic acid levels were detected by ELISA method. At 24 hours after injection, rat intestinal tissues were taken and observed pathologically under light microscopy, and their close connections were observed under transmission electron microscope. ZO-1 protein levels were detected by immunohistochemistry method.
    RESULTS AND CONCLUSION: Compared with the control group, the serum diamine oxidase, tumor necrosis factor α, and D-lactic acid levels were significantly lower in the experimental group at hours 6 and 24 after injection (P < 0.05). Intestinal necrosis, villous edema, intestinal congestion and inflammatory cell infiltration in the experimental group were milder than those in the control group. In addition, the ZO-1 protein expression in the experimental group was higher than that in the control group. Experimental results show that bone marrow mesenchymal stem cell transplantation into the intestinal mucosa can improve the intestinal mucosal permeability in rats with intestinal ischemia-reperfusion injury. 

     

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    Bone marrow mesenchymal stem cell transplantation combined with lithium chloride treatment for avascular necrosis of the femoral head in rabbits
    Li Wei, Lu Xiao-wei, Xian Cheng, Lao Shan
    2016, 20 (6):  868-875.  doi: 10.3969/j.issn.2095-4344.2016.06.016
    Abstract ( 289 )   PDF (703KB) ( 376 )   Save

    BACKGROUND: Lithium chloride can promote the proliferation and osteogenic capacity of bone marrow mesenchymal stem cells in the necrotic region after avascular necrosis of the femoral head, which has become an issue of concern.
    OBJECTIVE: To compare the advantages and disadvantages of bone marrow stem cell transplantation combined with lithium chloride in the treatment of rabbit femoral head necrosis.
    METHODS: Passage 2 bone marrow mesenchymal stem cells from 1-week-old New Zealand rabbits were cultured in 0, 5, 10, 20, 40 mmol/L lithium chloride. Forty-eight healthy adult New Zealand rabbits were selected to make femoral head necrosis models in the right femoral head using liquid nitrogen freezing method and then randomized into four groups: model group with no implantation; lithium chloride group given lithium chloride treatment at 3 days after modeling; cell transplantation group given gelatin sponge implantation and bone marrow mesenchymal stem cell suspension injection into the femoral head after modeling; combined group given bone marrow mesenchymal stem cell suspension injection and lithium chloride treatment. Intraperitoneal injection of lithium chloride (45.2 mg/kg) was given daily beginning at the postoperative 3rd day, and the treatment duration was 4 weeks.
    RESULTS AND CONCLUSION: Lithium chloride at 10 mmol/L had the maximum effect on the proliferation of rabbit bone marrow mesenchymal stem cells, and if the concentration of lithium chloride was > 10 mmol/L, the promotion role of lithium chloride began to decline. After combined treatment, the morphology of the femoral head was restored a little, with increased bone density and thickened trabecular bone; the level of β-catenin in the femoral head was significantly increased in the combined group compared with the cell transplantation group or the lithium chloride group. These findings show that bone marrow stem cell transplantation combined with lithium chloride treatment can promote the recovery from femoral head necrosis by increasing bone mass of the trabecular bone and bone density of the femoral head in the necrotic region. 

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    Early exercise training combined with neural stem cell transplantation improves hindlimb motor function after spinal cord injury in rats
    Wu Yu-jiang, Hou Ying-nuo, Zhang Zi-tan, Liu Zhong-po, Nie Zhi-hong, Fan Ge-lin
    2016, 20 (6):  876-882.  doi: 10.3969/j.issn.2095-4344.2016.06.017
    Abstract ( 259 )   PDF (542KB) ( 343 )   Save

    BACKGROUND: Studies have shown that neural stem cell transplantation combined with exercise training can promote the recovery of hindlimb motor function from spinal cord injury in rats, but its mechanism of action has not been fully elucidated.
    OBJECTIVE: To investigate the effects of early exercise training combined with neural stem cell transplantation on the recovery of hindlimb motor function in rats with spinal cord injury.
    METHODS: Sixty Sprague-Dawley rats with spinal cord injury were randomly divided into three groups: control group (n=20, given conventional treatment after injury), cell transplantation group (n=20, given neural stem cell transplantation after injury), experimental group, (n=20, given neural stem cell transplantation combined with early exercise training after injury). Recovery of the hindlimb motor function was assessed by Basso, Beattie and Bresnahan scale and inclined plane test before and at 1, 7, 14, 21 days after injury. Western blot assay was used to detect caspase-3 and myeloperoxidase expression. Hematoxylin-eosin staining was done at 21 days after injury to observe the structure changes of the injured spinal cord.
    RESULTS AND CONCLUSION: (1) Scores of Basso, Beattie and Bresnahan scale and inclined plane test were significantly better in the experimental group than the cell transplantation group followed by the control group (P < 0.05). (2) In the control group, the expression of caspase-3 and myeloperoxidase was significantly increased at 14 days after injury. In the cell transplantation, the expression of caspase-3 and myeloperoxidase was significantly higher than the experimental group (P < 0.05). (3) Pathological inflammation was reduced most in the experimental group followed by the cell transplantation group. In the experimental group, the structure of injured spinal cord was improved and became relatively clear and intact. These findings indicate that neural stem cell transplantation combined with early exercise training can effectively promote the recovery of hindlimb motor function from spinal cord injury in rats, by reducing the expression of caspase-3 and myeloperoxidase and alleviating secondary lesion of the spinal cord. 

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    Endothelial progenitor cell transplantation combined with early exercise training for spinal cord injury: improvement in hindlimb function and angiogenesis in the injured region
    Zhao Su-xiang, Hou Ying-nuo, Zhang Zi-tan, Liu Zhong-po, Nie Zhi-hong, Fan Ge-lin
    2016, 20 (6):  883-890.  doi: 10.3969/j.issn.2095-4344.2016.06.018
    Abstract ( 205 )   PDF (631KB) ( 341 )   Save

    BACKGROUND: Endothelial progenitor cells are widely used in the treatment of various vascular diseases, and early exercise training contributes to restore motor function after spinal cord injury. However, the therapeutic effects of endothelial progenitor cell transplantation or early exercise training alone are unfavorable.
    OBJECTIVE: To observe the influence of transplantation of endothelial progenitor cells combined with early exercise training on blood vessel regeneration and hind limb function in rats after spinal cord injury.
    METHODS: Eighty adult Sprague-Dawley rats were enrolled to establish spinal cord injury models using the modified Allen’s method, and then randomly divided into four groups. Rats were respectively given culture medium via the tail vein, injection of endothelial progenitor cells (3×106) via the tail vein, roller and treadmill trainings for 2 weeks, or injection of endothelial progenitor cells via the tail vein followed by 2 weeks of roller and treadmill trainings in the model, cell transplantation, exercise and combined groups.
    RESULTS AND CONCLUSION: At 2 weeks after transplantation, the hindlimb motor function of rats in the combined group was better than that in the cell transplantation group and exercise group, and moreover, the percentage of CM-Dil positive cells, the number of horseradish peroxidase-positive nerve fibers, capillary density and expression of vascular endothelial growth factor and brain-derived neurotrophic factor were also significantly higher in the combined group than the cell transplantation group and exercise group. These findings indicate that early exercise training has a neuroprotective role in spinal cord injury; endothelial progenitor cell transplantation combined with early exercise training can promote regeneration of synapses and blood vessels and improve hindlimb motor function of rats, probably by increasing expression levels of vascular endothelial growth factor and brain-derived neurotrophic factor.
     

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    Effect of basic fibroblast growth factor and insulin-like growth factor-1 on proliferation and collagen synthesis of bone marrow mesenchymal stem cells
    Xu Hai-long, Ding Yue, Xie Hong, Sun Xiao-ju, Xie Hui-xin
    2016, 20 (6):  891-897.  doi: 10.3969/j.issn.2095-4344.2016.06.019
    Abstract ( 260 )   PDF (504KB) ( 509 )   Save

    BACKGROUND: How to control the orderly formation of collage in skin repair and scarring process is worthy of attention.
    OBJECTIVE: To investigate the effect of basic fibroblast growth factor (bFGF) combined with insulin-like growth factor 1 (IGF-1) on the proliferation and collagen synthesis of rat bone marrow mesenchymal stem cells in vitro.
    METHODS: Rat bone marrow mesenchymal stem cells were isolated and cultured to induce adipogenic differentiation assessed by oil red O staining and osteogenic differentiation identified by alizarin red staining in vitro. Passage 3 cells were cultured in the medium containing bFGF, IGF-1, combination of them or the control fluid, respectively. MTT assay was used to detect cell proliferation at 12, 24, 48, 72 and 96 hours of culture. The expression of type I collagen and type III collagen were detected by RT-PCR and western blot after 10 days of incubation.
    RESULTS AND CONCLUSION: Compared with the control group, bFGF or IGF-1 alone significantly promoted the proliferation of bone marrow mesenchymal stem cells, and inhibited the expression of type I collagen and type III collagen. After combined use of bFGF and IGF-1, the proliferation of bone marrow mesenchymal stem cells was improved more significantly, and the expression of type I collagen and type III collagen returned to normal levels. These findings indicate that the combination of IGF-1 and bFGF can promote proliferation of bone marrow mesenchymal stem cells and restrain the expression of type I collagen and type III collagen, which may be helpful for control and repair of scar formation during wound healing. 

     

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    Biological properties of human periodontal ligament stem cells under inflammatory microenvironment
    Yuan Ping, Li Shu-hui, Zhao Lu, Yu Li, Zhou Chun-mei, Wu Pei-ling
    2016, 20 (6):  898-905.  doi: 10.3969/j.issn.2095-4344.2016.06.020
    Abstract ( 330 )   PDF (699KB) ( 810 )   Save
    BACKGROUND: The periodontal ligament stem cells can promote periodontal tissue regeneration, providing a new way for the treatment of periodontitis.
    OBJECTIVE: To observe the inflammatory microenvironment effects on the biological properties of periodontal ligament stem cells.
    METHODS: Periodontal ligament stem cells from healthy controls and patients with periodontitis were primarily cultured by tissue digestion method, purified using limited dilution method, and identified through detection of CD146 and STRO-1. Then, passage 3 cells were taken and denoted as normal control and inflammation groups followed by osteogenic induction.
    RESULTS AND CONCLUSION: Purified cells from two sources both expressed STRO-1 and CD146. Periodontal ligament stem cells in the inflammation group showed higher multiplication capacity, but the osteogenesis ability was lower compared with the normal control group. The expressions of Runx2 mRNA and Osterix mRNA were dropped significantly after the stimulus of tumor necrosis factor-α
    (P < 0.05), but the interleukin-1β and interleukin-6 did not have a significant impact. Tumor necrosis factor-α at 0.1 and 1 μg/L had no significant effects on the expression of Runx2 mRNA, but the expression of Runx2 mRNA was decreased significantly after treatment with 10 μg/L tumor necrosis factor-α (P < 0.05). It is confirmed that the molecular signaling mechanism inside the periodontal ligament stem cells is changed under inflammatory microenvironment, so that the differentiation capacity of cells from the inflammatory sources is lowered. Moreover, tumor necrosis factor-α is one of the key factors and its optimal concentration is 10 μg/L. 
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    Stem cell therapy for ischemic heart disease
    Miao Lei, Liu Pei-liang
    2016, 20 (6):  906-912.  doi: 10.3969/j.issn.2095-4344.2016.06.021
    Abstract ( 292 )   PDF (575KB) ( 566 )   Save

    BACKGROUND: Stem cell transplantation is a promising strategy for treatment of ischemic heart diseases, which has obtained great achievements in recent years.
    OBJECTIVE: To analyze the clinical trends of stem cell therapy for ischemic heart diseases reported from 2001 to 2015 in Medline, CNKI, Wanfang, ClinicalTrials.gov and Chinese Clinical Trial Register.
    METHODS: The relevant articles addressing stem cell therapy for ischemic heart diseases were retrieved using the keywords of “stem cell transplantation” and “ischemic heart disease” in Chinese and English in Medline, CNKI, Wanfang, ClinicalTrials.gov and Chinese Clinical Trial Register followed by statistical analysis. The retrieval time was from 2001 to 2015, including 2001-2005, 2006-2010 and 2011-2015.
    RESULTS AND CONCLUSION: In Medline database, there were 219 clinical studies about stem cell therapy for ischemic heart disease published from 2001 to 2015, and the number of retrieved articles was most from 2006 to 2010, and decreased from 2011 to 2015. In CNKI and Wanfang databases, the number of relevant articles which had a similar trend with that in the Medline database decreased remarkably after 2011. In ClinicalTrials.gov, there were 63 clinical trials about stem cell therapy for ischemic heart disease, most of which were registered from 2006 to 2015 and came from Europe and North America. In Chinese Clinical Trial Register, there was no clinical trial about stem cell therapy for ischemic heart disease, which may result from the strict management in China. 

     

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