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04 March 2012, Volume 16 Issue 10 Previous Issue    Next Issue

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Distribution of green fluorescent protein transgenic mouse bone marrow mesenchymal stem cells in various organs of rats with severe acute pancreatitis Jin Shan-feng, Chen Zhi-yao, Huang He-guang 2012, 16 (10):  1711-1715.  doi: 10.3969/j.issn.1673-8225.2012.10.001 Abstract ( 217 )   PDF (537KB) ( 393 )   Save BACKGROUND: Green fluorescent protein transgenic mouse bone marrow mesenchymal stem cells are transplanted into rats with severe acute pancreatitis for easy in vitro tracing in animals because of the characteristic of carrying fluorescence. OBJECTIVE: To investigate the distribution of green fluorescent protein transgenic mouse bone marrow mesenchymal stem cells in various organs of rats with severe acute pancreatitis. METHODS: Green fluorescent protein transgenic mouse bone marrow mesenchymal stem cells were isolated directly by adherence. After reaching 90% confluency, cells were digested, passaged, and proliferated. After three passages, cell immunophenotype of CD29+, CD90+, CD34-, CD45- were determined. Sprague-Dawley rat model of severe acute pancreatitis were established by retrograde injection of 5% sodium taurocholate into the pancreatic duct under the microscope. 1 hour later, cells at a density of 2×106/rat were injected into rats with severe acute pancreatitis via tail vein. At 6, 12, 24 hours, the liver, kidney, brain, lung, pancreas, and intestine were harvested for pathological examination. The pathological changes of the pancreas and the distribution of green fluorescent protein transgenic mouse bone marrow mesenchymal stem cells in various organs of rats with severe acute pancreatitis as well as gray scale values were determined. RESULTS AND CONCLUSION: The pancreas was more seriously destroyed with the time and it was most seriously destroyed at 24 hours. 91.1% of green fluorescent protein transgenic mouse mesenchymal stem cells were positive for CD29, 93.5% positive for CD90,0.82% positive for CD34, and 2.22% positive for CD45. After injection of stem cells, green fluorescence was observed in each organ and enhanced with time. The gray scale of green fluorescence was highest in the kidney tissue and lowest in the brain tissue. These findings suggest that the transplanted bone marrow mesenchymal stem cells can be stably distributed in each organ of rats with severe acute pancreatitis. Related Articles | Metrics

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In vivo homing of chloromethyl-benzamidodialkylcarbocyanine-labeled bone marrow mesenchymal stem cells after transplantation into a mouse model of immune-mediated bone marrow failure Wang Yang1, 2, Xiao Yang2, Li Li2, Jiang Zu-jun2, Li Yong-hua2, Pang Yan2, Song Jia-yin2 2012, 16 (10):  1716-1720.  doi: 10.3969/j.issn.1673-8225.2012.10.002 Abstract ( 260 )   PDF (524KB) ( 278 )   Save BACKGROUND: Aplastic anemia (AA) is characterized by pancytopenia and bone marrow (BM) hypoplasia. In most cases, AA is an immune-mediated disease, with active destruction of hematopoietic stem and progenitor cells by T lymphocytes. Bone marrow mesenchymal stem cells (BMSCs) can support hematopoietic cells and have immunomodulatory effects. OBJECTIVE: To observe the homing of BMSCs transferred into mouse model of bone marrow failure. METHODS: BALB/C mice were randomly divided into normal control group, AA group and bone marrow mesenchymal stem cell transplantation group. On day 6, chloromethyl-benzamidodialkylcarbocyanine (CM-Dil)-labeled BALB/c mouse bone marrow mesenchymal stem cells were transplanted into mouse model of bone marrow failure via tail vein. The dynamic distribution of CM-Dil-labeled cells in different tissues was observed by flow cytometry and fluorescence microscope. On day 14 after bone marrow failure induction, the average survival time, peripheral blood hemogram and the morphological features of bone marrow of mice in each group were determined. RESULTS AND CONCLUSION: At 24 hours after BMSC transplantation via tail vein, CM-Dil-labeled positive cells were observed in recipients’ bone marrow, and the CM-Dil-labeled positive cells increased at 72 hours (P < 0.05). The white blood cells, hemoglobin, platelets and bone marrow mononuclear cells of the dying or executed mice in the transplantation group were significantly higher than those in the aplastic anemia group (P < 0.01). The average survival time of mice in aplastic anemia group was significantly shorter than that in the BMSC transplantation group (P < 0.05). These findings suggest that the transplanted BMSCs can home into the bone marrow of bone marrow failure model, promote the recovery of hematopoietic function, alleviate the degree of bone marrow failure, and significantly prolong survival time.    Related Articles | Metrics

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Effects of 17 beta-estradiol on osteogenic differentiation of rat primary cultured bone marrow mesenchymal stem cells Han Na1, Kou Yu-hui2, Wang Tian-bing2, Yin Xiao-feng2, He Pei-ying1, Zhang Pei-xun2 2012, 16 (10):  1721-1724.  doi: 10.3969/j.issn.1673-8225.2012.10.003 Abstract ( 290 )   PDF (387KB) ( 326 )   Save BACKGROUND: The estrogen receptor exists in bone marrow mesenchymal stem cells and estrogen promotes bone formation by regulating the proliferation and differentiation of bone marrow mesenchymal stem cells. OBJECTIVE: To explore the effects of 17β-estradiol on the osteogenic differentiation of bone marrow mesenchymal stem cells (MSCs) and the underlying mechanism. METHODS: MSCs were separated, purified, cultured and expanded. Different concentrations of 17β-estradiol was administrated to MSCs for some period. The level of type I collagen in culture medium was detected by Elisa method, and the expression of Runx2 mRNA and protein was analyzed by RT-PCR or Western blotting methods, respectively. RESULTS: Compared with the control group, type I collagen expression in the MSCs was significantly increased on the 5th day after administration of 0.001, 0.01, 0.1 nmol/L 17β estradiol (P < 0.05) and it maintained high expression on the 7th day (P < 0.05). Runx2 mRNA expression increased in a dose-dependent manner on the 5th and 7th days after administration of 17 β-estradiol (P < 0.05). These findings suggest that 17β-estradiol can induce the osteogenic differentiation of MSCs, which occurs possibly by upregulation of Runx2 expression. Related Articles | Metrics

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Effects of low frequency vibration on osteogenesis of bone marrow stromal cells and receptor activator of nuclear factor-kappa B/receptor activator of nuclear factor kappa-B ligand/osteoprotegerin pathway Du Guang-yu, Zhao Wen-zhi, He Sheng-wei, Mi Li-dong, Zhang Lu, Sun Chuan-xiu, Sun Xue-gang 2012, 16 (10):  1725-1728.  doi: 10.3969/j.issn.1673-8225.2012.10.004 Abstract ( 256 )   PDF (502KB) ( 322 )   Save BACKGROUND: There have been few studies regarding low frequency vibration stimulating osteogenic differentiation of bone marrow stromal cells in vivo. OBJECTIVE: To investigate the effects of different frequencies of vibration on receptor activator of nuclear factor-kappa B (RANK)/ receptor activator of nuclear factor kappa-B ligand (RANKL)/osteoprotegerin (OPG) pathway and preliminarily investigate the underlying mechanism.  METHODS: Eighty New Zealand rabbits of bone defects were established. After transplantation of complex of New Zealand bone marrow stromal stem cells and decalcified bone matrix into bone defect region, rat models were randomly divided into a control group and four vibration groups according to different vibration frequencies (12.5, 25, 50, 100 Hz). The vibration group received different frequencies of vibration for 5 weeks from day 7. After vibration, OPG mRNA and RANKL mRNA expression was detected. RESULTS AND CONCLUSION: Compared with the control group, OPG and RANKL mRNA expression was significantly up regulated in each vibration group (P < 0.05), in particular in 25 and 50 Hz vibration groups (P < 0.01), but the expression was significantly down regulated in the 100 Hz vibration group (P < 0.05). This suggests that vibration at a certain frequency can stimulate bone marrow stromal cells to repair bone defects, which occurs possibly because vibration can promote the up regulation of OPG mRNA expression, with the ideal vibration frequency of 25 and 50 Hz. Related Articles | Metrics

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Application of myocardial tissue lysates on the differentiation of bone marrow mesenchymal stem cells into cardiac tissue-like structure  Zhao Wen-jing, Li Qin, Wu Zhuo, Chen Wei-ping 2012, 16 (10):  1729-1732.  doi: 10.3969/j.issn.1673-8225.2012.10.005 Abstract ( 258 )   PDF (432KB) ( 342 )   Save BACKGROUND: In vivo experiment has shown that the microenvironment of bone marrow mesenchymal stem cells (BMSCs) plays an important role on induced differentiation. OBJECTIVE: To in vitro induce the BMSCs to differentiate into cardiac tissue-like structure. METHODS: Changes of morphological characteristics of the third generation of BMSCs that cultured with myocardial tissue lysates for 4 weeks were observed under inverted microscope. The tissue was collected and preformed with hematoxylin-eosin staining, and the ultrstructure of the induced cells was observed by electron microscope. Immunohistochemical staining was preformed using vimentin and Ⅷ factor-related antigen. RESULTS AND CONCLUSION: Myocardial tissue lysates were cultured for 4 weeks. At the 12th day, some “bamboo-like” cells were fused to form multinucleated myotube-like structure. From the 14th day, rhythmic contraction could be observed with the beating frequency about 90-110/min; sarcomere-like structure of filaments could be seen under transimission electron microscope and some muscle cells had the function of automatic pulsating. Hematoxylin-eosin staining showed a large number of bundle-like structures. Myocardial tissue lysates can be used as simulated cardiac tissue microenvironment inducer to promote the BMSCs to differentiate into myocardial cells with typical morphological features and the corresponding functions, and induce part of the BMSCs to differentiate into connective tissue cells, endothelial like cells, and form cardiac tissue-like structure together with cardiac cells. BMSCs have the multiple differentiation potential, forming a variety of cells with similar functions and forming the tissue-like structure trend in a certain microenvironment. Related Articles | Metrics

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Separation, culture and identification of rat bone marrow-derived endothelial progenitor cells Wang Li1, Zhang Hui-feng2, Yuan Hui-juan2, Ma Yue-hua2, Wang Yan-fang2, Hu Zi-ying2, Su Yong2, Zhao Zhi-gang2 2012, 16 (10):  1733-1736.  doi: 10.3969/j.issn.1673-8225.2012.10.006 Abstract ( 221 )   PDF (426KB) ( 352 )   Save BACKGROUND: Endothelial progenitor cells (EPCs) are involved in angiogenesis, but its isolation, culture and identification methods are not uniform currently. OBJECTIVE: To explore the methods of separation, culture and identification of rat bone marrow-derived EPCs. METHODS: EPCs were cultured using density gradient centrifugation and differential velocity adherent methods, growth and morphology changes of cells were observed under inverted microscope. The expression of cell surface antigen CD34 and CD133 were detected by using Dil labeled acetylated low-density lipoprotein (Dil-ac-LDL) and FITC-labeled Ulex europaeus agglutinin 1 (FITC-UEA-1) double dyeing and flow cytometry.  RESULTS AND CONCLUSION: For the first four days, cell proliferation was not obvious, at the 5-10 days, cell proliferation was obvious, and colony-forming unit (CFU) and lineage structure could be seen. At the 7th day, EPCs could swallow the function of DiL-ac-LDL and FITC-UEA-1. At the 10th day, in vitro flow cytometry detection showed that the CD133+ cells were accounted for 19.2%, CD34+ cells were accounted for 28.7%, CD34+/CD133+cells were accounted for 19.1%. Using density gradient centrifugation and differential velocity adherent methods can separate and culture bone marrow derived EPCs. Related Articles | Metrics

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In vitro induction of human bone marrow mesenchymal stem cells to differentiate into type Ⅱalveolar epithelial cells Chen Yin1, 2, Ma Nan2, Mei Ju2, Xiao Hai-bo2, Lu Shan-wei1, Xu Huai-Yang1, Zhong Hong1, 2 2012, 16 (10):  1737-1741.  doi: 10.3969/j.issn.1673-8225.2012.10.007 Abstract ( 278 )   PDF (515KB) ( 437 )   Save BACKGROUND: Alveolar epithelial type Ⅱ (AT Ⅱ) has been demonstrated to directly participate in liver recovery and can also alleviate the severity of pulmonary fibrosis in a rat model of pulmonary fibrosis. Embryonic stem cells (ESCs) can differentiate into AT Ⅱ in vitro, but the application of ESCs is confined by many factors. OBJECTIVE: To investigate the method and transformation rate of bone marrow mesenchymalstem cells (BMSCs) differentiation into AT Ⅱ. METHODS: The BMSCs were isolated from human bone marrow cells. BMSCs were co-cultured with human embryonic lung mesenchymal cells (MRC-5) in serum-free small airway growth medium (SAGM) and modified SAGM. The morphology of BMSCs was observed, and the expression of surfactant protein C (SPC) mRNA was detected by RT-PCR. In addition, the expression of SPC was also detected by immunofluorescence. RESULTS AND CONCLUSION: After the BMSCs were co-cultured with MRC-5 for 10 days, some of BMSCs began to turn into epithelioid cells in morphous. After 15 days, the expression of SPC mRNA was detected, but it was not increased along with the prolongation of coculture days. Compared with SAGM, modified SAGM could significantly increase the expression of SPC mRNA (P < 0.05). BMSCs could differentiate into AT Ⅱ in SAGM or modified SAGM when co-culture with MRC-5 in vitro at low transformation rate. Positive rate of SPC was only (3.15±0.69)%. Related Articles | Metrics

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Single-cell clonal culture and identification of human bone marrow stromal stem cells   Teng Yong1, Xu Jian-li2, Li Xu-sheng1, Hu Yun-yu3, Wang Zhen3 2012, 16 (10):  1742-1747.  doi: 10.3969/j.issn.1673-8225.2012.10.008 Abstract ( 331 )   PDF (542KB) ( 300 )   Save BACKGROUND: Bone marrow stromal stem cells lack of specific surface marker and their identification challenges scholars all the time. OBJECTIVE: To explore the culture condition for clonal isolation of human bone marrow stromal stem cells (BMSCs) and identify their surface markers and differentiation potentials. METHODS: Human bone marrow was taken and BMSCs were isolated using density gradient centrifugation. Clone-like cells were selected by single cell limiting dilution. The surface antigens of the cloned BMSCs were analyzed by flow cytometry and immunocytochemistry. Multi-differentiation capacities were evaluated by inducing BMSCs into osteoblasts, chondrocytes and cadiocytes. RT-PCR and immunocytochemistry were applied to detect the three mutli-differentiations. RESULTS AND CONCLUSION: Single cell-derived BMSCs could be expanded to passage 28 in vitro which still maintaued active proliferation ability.The expanded BMSCs expressed CD29, CD44, CD106, but not CD14, CD34, CD45, HLA-DR. They could be induced to differentiate into osteoblasts, chondrocytes and cadiocytes. These findings suggest that cloned BMSCs can maintain the phenotypes of stem cells during in vitro culture. Related Articles | Metrics

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Bone marrow-derived mesenchymal stem cells for ventricular remodeling following acute myocardial infarction  Yu Gui-Ping1, Shen Zhen-ya2, Yu Yun-sheng2, Guo Shi-qiang2, Chen Yi-huan2, Hu Yan-qiu2 2012, 16 (10):  1748-1751.  doi: 10.3969/j.issn.1673-8225.2012.10.009 Abstract ( 215 )   PDF (349KB) ( 301 )   Save BACKGROUND: Animal experiments and preliminary clinical studies have demonstrated that transplantation of stem cells can replace necrotic myocardial cells, increase the number of myocardial cells with functions and improve cardiac function, thereby providing a brand-new pathway for treatment of myocardial infarction. OBJECTIVE: To investigate the effects of transplantation of autologous bone marrow mesenchymal stem cells and bone marrow mobilization after myocardial infarction on cardiac functions. METHODS: 30 mL bone marrow was taken from pig anterior superior iliac spine for harvesting bone marrow mesenchymal stem cells. Fifteen pigs were assigned to three groups. In the model group, only myocardial infarction was induced. In the stem cell transplantation group, at 3 hours after myocardial infarction induction, bone marrow mesenchymal stem cells were transfused via coronary vein. In the stem cell mobilization group, at 3 hours after myocardial infarction induction, granulocyte colony-stimulating factor (150 μg/kg per day) was administered for a total of 5 successive days. RESULTS AND CONCLUSION: In the stem cell mobilization group and stem cell transplantation group, left ventricular end systolic diameter and left ventricular end-diastolic diameter were slightly, but not significantly, decreased, and ejection fraction was slightly, but not significantly, increased (P > 0.05) at 8 weeks after myocardial infarction compared with before infarction. In the stem cell mobilization group and stem cell transplantation group, vascular endothelial growth factor level was significantly increased at 8 weeks after myocardial infarction than before infarction (P < 0.05). At 8 weeks after myocardial infarction, capillary density at the infarcted zone was significantly greater in the stem cell mobilization group and stem cell transplantation group than in the model group (P < 0.05). These findings suggest that after myocardial infarction, autologous bone marrow mesenchymal stem cell transplantation and bone marrow mobilization can significantly improve cardiac function, but the precise efficacy requires further investigation via large-size sample.  Related Articles | Metrics

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富集骨髓间充质干细胞用于兔腰椎横突间的融合**☆ Sun Wei1, Zhao Jie1, Guo Kai-jin2, Yuan Feng2 2012, 16 (10):  1752-1755.  doi: 3969/j.issn.1673-8225.2012.10.010 Abstract ( 214 )   PDF (366KB) ( 252 )   Save BACKGROUND: Nano-hydroxyapatite/collagen has good cellular compatibility. It can create three-dimensional environment for cells growth and be well used as bone carrier material based on its nano-porous structure. OBJECTIVE: To explore the feasibility of the enriched bone marrow stromal stem cells (BMSCs) composite with nano-hydroxyapatite/collagen (nHAC) for lumbar intertransverse spinal fusion of rabbits. METHODS: Twenty New Zealand rabbits were divided into the cells group (n=10) and the autograft group (n=10) according to their different implants in bilateral L5-L6 intertransverse process. Drawing materials was performed 8 weeks after implantation. Then the technique detection and histological observation were performed in order to estimate the lumbar fusion. RESULTS AND CONCLUSIONS: The fusion rates in the cells group and the autograft group were 70% (7/10) and 80% (8/10) respectively 8 weeks after implantation, and had no significant difference (P > 0.05). Histological observation showed that there was more newly formed bone trabeculae growth inside the nHAC of the cells group. A great amount of new bone tissues and linked bone trabeculae were seen in intertransverse process of the autograft group. This study showed that enriched BMSCs composite with nHAC can be used as a substitute for autograft in spine fusion, and is similar to autograft in spinal fusion. Related Articles | Metrics

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Simulation of stem cell microenvironment in vitro to culture and expand cord and umbilical cord mesenchymal stem cells Qiu Yun, Zheng Qing, Xiao Shu-dong, Wang Zheng 2012, 16 (10):  1756-1760.  doi: 10.3969/j.issn.1673-8225.2012.10.011 Abstract ( 300 )   PDF (519KB) ( 355 )   Save BACKGROUND: The future of cell therapy requires high purity of mesenchymal stem cells (MSCs) in order to clearly determine the therapeutic dose and dose-response relationship. Establishing a robust amplification system in vitro to obtain the therapeutic dose of stem cells while preserving the characteristics of stem cells is the urgent of clinical laboratories. OBJECTIVE: To simulate a microenvironment for stem cells in vitro to harvest and expand umbilical cord blood-derived and cord-derived MSCs and to compare with conventional two-dimensional culture system. METHODS: The umbilical cord blood-derived and cord-derived MSCs were inoculated in the conventional two-dimensional plastic culture system and extracellular matrix, respectively. The two amplification systems for in vitro large-scale expansion of umbilical cord blood-derived and cord-derived MSCs were evaluated from the points of cell counts and surface markers. RESULTS AND CONCLUSION: The production of umbilical cord blood-derived and cord-derived MSCs in the bone marrow-derived extracellular matrix culture system was 4-6 times of that in the conventional two-dimensional culture system. FACS analysis showed that extracellular matrix system better maintained the surface marker of stem cells. Therefore, such a three-dimensional culture system is much closer to physical environment than the two-dimensional culture system. The established bone marrow-derived extracellular matrix culture system can maintain the characteristics of mesenchymal stem cells, which provides access to more homogeneous MSCs of high activity within a short time period. Related Articles | Metrics

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Effect of recombinant human erythropoietin on differentiation of myocardial precusor cells from amniotic fluid stem cells of pregnancy women Liu Yan1, Li Fa-Qi1, Zhang Bin2, Luo Jian-Ping3 2012, 16 (10):  1761-1764.  doi: 10.3969/j.issn.1673-8225.2012.10.012 Abstract ( 222 )   PDF (406KB) ( 269 )   Save BACKGROUND: Studies have shown that amniotic fluid stem cells of pregnancy women can differentiate into myocardial precursor cells. OBJECTIVE: To study the effect of recombinant human erythropoietin (rhEPO) at different concentrations on differentiation of myocardial precusor cells from amniotic fluid stem cells of pregnancy women. METHODS: The differentiation of myocardial precursor cells from amniotic fluid stem cells of pregnancy women was induced by adding conditional culture medium containing 0, 1.0, 5.0, 10.0, 20.0 U/mL rhEPO. After 24 hours, conditional culture medium was replaced by complete medium. RESULTS AND CONCLUSION: During 14 days of induced differentiation, under the inverted microscope, cells gradually shortened and widened from original shuttle appearance. Adjacent cells fused together, similar to myotube-like structure. The differentiation rate of myocardial precursor cells was highest after 5.0 U/mL rhEPO addition. RT-PCR results showed that rhEPO particularly at the concentration of 5.0 U/mL could promote Nkx-2.5 and GATA-4 mRNA expression. These findings suggest that exogenous rhEPO can promote the differentiation of myocardial precursor cells from amniotic fluid stem cells of pregnancy women dose-dependently. Related Articles | Metrics

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Human umbilical cord mesenchymal stem cells influence the proliferation and apoptosis of endometrial cells  Xu Li-nan1, Xu Bing-nan2, Chen Shu-qin1, He Ke1, Cai Jian1 2012, 16 (10):  1765-1768.  doi: 10.3969/j.issn.1673-8225.2012.10.013 Abstract ( 250 )   PDF (321KB) ( 344 )   Save BACKGROUND: Human umbilical cord mesenchymal stem cells (hUCMSCs) can regulate cellular biological activities by paracrine mechanism. OBJECTIVE: To investigate the effects of hUCMSCs on the proliferation and apoptosis of endometrial cells. METHODS: Endometrial cells and hUCMSCs were cultured in vitro. hUCMSCs and endometrial cells were co-cultured on the Transwell co-culture plate at a proportion of 1×105/1×105 to establish upper and lower double-cell non-contact co-culture system, serving as the experimental group. Endometrial cells (1×105 cells/well) were separately cultured as control group. RESULTS AND CONCLUSION: MTT assay results showed that cell proliferation in the experimental group was significantly inhibited in a time-dependent manner compared with the control group (P < 0.05). Flow cytometry results showed that apoptotic cells were significantly more in the experimental group than in the control group (P < 0.05), and endometrial cells entering into S stage from G1 stage were reduced. RT-PCR results showed that PTEN mRNA expression was significantly higher in the experimental group than in the control group. These findings suggest that hUCMSCs inhibit the in vitro proliferation of human endometrial cells and promote their apoptosis by paracrine mechanism, which occurs possibly by upregulation of PTEN mRNA expression. Related Articles | Metrics

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Human umbilical cord stromal stem cells-conditioned medium down-regulated the expression of transforming growth factor beta in rat hepatic stellate cells Li Ming-liang, Yao Zhi-cheng, Zhong Yue-si, Lin Nan, Xu Rui-yun, Deng Mei-hai 2012, 16 (10):  1769-1772.  doi: 10.3969/j.issn.1673-8225.2012.10.014 Abstract ( 280 )   PDF (387KB) ( 264 )   Save BACKGROUND: Human umbilical cord stromal stem cells are expected to become a new seed cell in the treatment of live cirrhosis. However, few studies of in vitro experiment have been reported. OBJECTIVE: To explore the effect of human umbilical cord stromal stem cells-conditioned medium (USSC-CM) on the biological activity of fibrosis in rat hepatic stellate cells (HSCs), and the acting target. METHODS: HSCs were seeded in the six orifice plate in proportion (1.0×105 /mL), then cultured without serum for 24 hours; 2 mL USSC-CM were added to treat HSCs as experimental group; while HSCs were treated with equivalent fresh medium as control group. The expression of transforming growth factor β1 (TGF-β1), collagen-α1 and TIMP-2 mRNA in HSCs were detected by RT-PCR. The expression of TGF-β1 protein was determined by Western blot. RESULTS AND CONCLUTION: The expression of TGF-β1, collagen-α1 and TIMP-2 mRNA in HSCs were down-regulated in experimental group compared to control group, and the tendency enhanced with time (USSC-CM treated 72 h > USSC-CM treated 48 h > control group). And the expression of TGFβ1 protein in HSCs were down-regulated in experimental group compared to control group and showed time dependent manner (control group > USSC-CM treated 48 h group > USSC-CM treated 72 h group, P < 0.05). Human umbilical cord stromal stem cells can inhibit biological activity of HSCs. And it may carry its point by focusing on the acting target related to TGF-β together with downstream production. Related Articles | Metrics

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Biological characteristics, osteogenic and adipogenic differentiation of mesenchymal stem cells from human umbilical cord blood*★ Zhang Cai-long1, Na Na2, Zhang Ji-hua3, Sun Kang1, Tian Shao-qi1, Xia Chang-suo1 2012, 16 (10):  1773-1779.  doi: 10.3969/j.issn.1673-8225.2012.10.015 Abstract ( 225 )   PDF (380KB) ( 214 )   Save BACKGROUND: Up to now, there are few reports addressing the biological properties and differentiation potential of umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs). OBJECTIVE: To observe the biological characteristics, osteogenic and adipogenic differentiation potential of UCB-MSCs. METHODS: MSCs were harvested and cultured from UCB at various gestational ages (GA). Harvested UCB-MSCs were cultured primarily and subcultured, and then induced to differentiate into osteoblasts and adipocytes. RESULTS AND CONCLUSION: Under the inverted phase contrast microscope, UCB-MSCs adhered to the wall, showing fibroblast like morphology and whirlpool like growth alignment. Observation of the ultramicrostructure under transmission electron microscope showed that UCB-MSCs had a big cell nucleus, fewer cellular organelles and big karyoplasmic ratio. All of the growth curves of primary and passaged UCB-MSCs presented S-shaped. The 3rd and 5th passages of MSCs showed the strongest proliferation activity. The count of colony forming unit fibroblasts varied with GA, significant difference was found among the three GA groups (P < 0.05), and the lower GA group had a higher count of colony forming unit fibroblasts than that in the older GA group. Flow cytometry showed that these cells expressed CD29, CD44 and CD90 positively, but they failed to express hematopoiesis related molecules such as CD34 and CD45. When the MSCs were induced to osteogenic and adipogenic differentiation for 3 weeks, strong expression of alkaline phosphatase was found and the formation of a mineral extracellular matrix was detected by alizarin red staining were detected; and neutral lipid vacuoles were detected by oil red O staining. UCB-MSCs have similar morphological and biological characteristics and cell surface molecule markers with MSCs derived from bone marrow, both of which have great capability of proliferation and regeneration. UCB-MSCs can be induced to osteoblasts and adipocytes in a suitable condition in vitro. Related Articles | Metrics

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Biological characteristics and optimal strategy of human amniotic fluid-derived mesenchymal stem cells cultured in vitro    Wang Pei-pei1, Zheng Yu-bao1, Peng Liang1, Yu Jun2, Gao Zhi-liang1 2012, 16 (10):  1780-1785.  doi: 10.3969/j.issn.1673-8225.2012.10.016 Abstract ( 292 )   PDF (511KB) ( 344 )   Save BACKGROUND: At present, seed sources of stem cells engineering are diverse, and mesenchymal stem cells can be isolated from exfoliated cells in fetal amniotic fluid. OBJECTIVE: To investigate the effects of the media with different elements on human amniotic fluid-derived mesenchymal stem cells (h-AFMSCs) cultured in vitro, and to analyze the biological characterization of h-AFMSCs at optimum medium cultured in vitro in order to optimize the culture strategy of h-AFMSCs. METHODS: Amniotic fluid was extracted from pregnant women during 16 to 24 weeks of gestation, and then was used for primary culture and subculture. h-AFMSCs were cultured in media containing six different elements. RESULTS AND CONCLUSION: h-AFMSCs were isolated from amniotic fluid in mid-trimester. The proliferative ability of h-AFMSCs was improved to a maximal extent in the mixed medium of low glucose Dulbecco's modified eagle medium containing 10% fetal bovine serum and MESEN PRO medium at a mixed ratio of 1:1. h-AFMSCs highly expressed CD29, CD73, CD90, CD105, CD166 and lowly expressed CD14, CD34, CD45, HLA-DR. Stem cells gene OCT-4 and Naong highly expressed in h-AFMSCs cultured in vitro. h-AFMSCs showed a higher osteogenic and adipogenic differentiation potential, and they could inhibit the proliferation of lymphocytes in vitro. It is indicated that mid-trimester amniotic fluid is a good cell source of h-AFMSCs with lower immunogenicity. Related Articles | Metrics

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Preparation of CF-1 mouse embryonic fibroblast feeder layers Lu Hai1, Zhang Zhi1, Zhao Yi-chao1, Rao Xiao-hui1,Jiang Jin-qun2, Meng Bin3, Ai Min4, Pan Ming-xin1, Gao Yi1 2012, 16 (10):  1786-1790.  doi: 10.3969/j.issn.1673-8225.2012.10.017 Abstract ( 382 )   PDF (356KB) ( 344 )   Save BACKGROUND: Mouse embryo fibroblasts as the feeder layer for growth of stem cells exhibit better effects than the circumstance without feeder layer because they can secrete some factors which can promote the growth of stem cells and inhibit the differentiation of stem cells. OBJECTIVE: To establish the optimal method for isolation and culture of CF-1 mouse embryo fibroblast feeder layer and investigate the optimal concentration of mitomycin C in inhibiting the proliferation of fibroblasts and the processing time. METHODS: Fetal mice were taken from CF-1 mice at pregnancy days 10-15. Primary fibroblasts were isolated and treated with different concentrations (5, 10, 15, 20 mg/L) of mitomycin C for different time periods (1, 1.5, 2, 2.5, 3 hours) to prepare feeder layers. Cell proliferation was observed. Human induced pluripotent stem cells or embryonic stem cells were cultured on mitomycin C-treated feeder layer and the growth of cell colonies was observed. RESULTS AND CONCLUSION: The optimal fetal age for preparing CF-1 mouse embryo fibroblast feeder layer was 13.0-14.0 days. Treatment with 10 mg mitomycin C for 2.5 hours showed optimal effects on inhibiting the proliferation of CF-1 mouse embryo fibroblasts and fibroblast feeder layer could maintain 5-7 days. Induced pluripotent stem cells and embryonic stem cells can develop into typical “bird nest”-shaped colony of stem cells after treatment with 10 mg/L mitomycin C for 2.5 hours. Related Articles | Metrics

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In vitro induction method for the differentiation of human peripheral blood mononuclear cells into hepatocyte-like cells  Wang Kai, Zhu Ying, Liu Jing, Zhao Gang 2012, 16 (10):  1791-1794.  doi: 10.3969/j.issn.1673-8225.2012.10.018 Abstract ( 245 )   PDF (364KB) ( 269 )   Save BACKGROUND: Peripheral blood mononuclear cells can differentiate into hepatocyte-like cells in vitro. OBJECTIVE: To explore the induced method that peripheral blood mononuclear cells can be differentiated to hepatocyte-like cells in vitro. METHODS: Peripheral blood mononuclear cells were isolated from patients by using density gradient centrifugation and cells adherent method, and cultured in the culture medium containing recombinant human macrophage colony stimulating factor (M-CSF), interleukin 3 (IL-3) and β-Mercaptoethanol for 6 days. The purified cells were cultured with hepatocyte growth factor and fibroblast growth factor 4 to accelerate induction for 14 days (experimental group). Non-induced mononuclear cells and L02 liver cell line were used as controls. RESULTS AND CONCLUSIONS: Freshly isolated peripheral blood mononuclear cells were small round, uniform size, strong refraction. After induction with M-CSF, IL-3 and β -Mercaptoethanol for 6 days, the part of cells grew into fibroblasts; and after induction with HGF and FGF-4 for 14 days, the cell morphology was similar to liver cells, and the cells expressed albumin, alpha-fetoprotein, cytokeratin 18. Peripheral blood mononuclear cells can be transformed into liver cells in vitro under certain induced conditions. Related Articles | Metrics

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Effect of uric acid on proliferation of human placenta-derived mesenchymal stem cells Cui Jing1, Yang Nai-long1, Wang Jing-yi2 2012, 16 (10):  1795-1798.  doi: 10.3969/j.issn.1673-8225.2012.10.019 Abstract ( 232 )   PDF (333KB) ( 362 )   Save BACKGROUND: Uric acid (UA) plays different roles on different cells. But, what is the effect of UA on the proliferation of human placenta-derived mesenchymal stem cells (hPMSCs), promoting or suppression? 0BJECTIVES: To observe the effect of UA on proliferation of hPMSCs, and to explore the relationship between time-effect and dose-effect. METHODS: The hPMSCs cultured in vitro were used and divided into 6 groups: the control group and the UA group which contained 0.1, 0.2, 0.4, 0.6, 0.8 mmol/L UA. RESULTS AND CONCLUSION: MTT assay showed that the number of hPMSCs in 0.1, 0.2, 0.4, 0.6, 0.8 mmol/L UA group was more than that in the control group at the same time, especially in 0.8 mmol/L UA group; the number of hPMSCs in same concentration of UA was increasing following the time (time﹤5 d), and reached to peak at the 5th day. UA could accelerate the proliferation of hPMSCs and showed a time- and dose-dependent manner, and the effect of 0.8 mmol/L UA on the proliferation capacity of hPMSCs was stronger than others at the 5th day. Related Articles | Metrics

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In vitro induced differentiation of pancreatic stem cells into islet-like cell clusters Cen Yan-hui, Xie Xiao-xun, Chen Wei-ping, He Guo-zhen, Huang Bo, Guo Wen-wen, Xiao Yan-ziCen Yan-hui, Xie Xiao-xun, Chen Wei-ping, He Guo-zhen, Huang Bo, Guo Wen-wen, Xiao Yan-zi 2012, 16 (10):  1799-1802.  doi: 10.3969/j.issn.1673-8225.2012.10.020 Abstract ( 239 )   PDF (310KB) ( 312 )   Save 背景：目前由于胰岛来源匮乏，使得胰岛细胞移植治疗糖尿病无法满足临床需求，故体外将胰腺干细胞诱导分化为胰岛成为研究焦点。 目的：于体外将小鼠胰腺干细胞诱导成胰岛样细胞团并对其进行相关检测，探寻一种胰腺干细胞体外诱导分化成胰岛及鉴定的技术和方法。 方法：体外获得纯化的小鼠胰腺干细胞，采用联合诱导剂对其进行成胰岛方向的诱导分化，并对诱导形成的胰岛样细胞团进行形态学观察、双硫腙染色、RT-PCR和Western blot检测。 结果与结论：实验通过细胞形态学和细胞生长特性的观察以及免疫细胞化学染色证实体外成功获得了小鼠胰腺干细胞，采用联合诱导剂将其诱导成胰岛样结构，呈球形，以较细长的蒂部与瓶底连接，双硫腙染色将其染成铁红色。RT-PCR和Western blot 法可分别检测到胰岛样细胞团的胰岛素mRNA和胰岛素蛋白。结果证实小鼠胰腺干细胞可体外诱导分化成含β细胞的胰岛样细胞团。 关键词：胰腺干细胞；诱导；胰岛样细胞团；干细胞培养；小鼠 doi:10.3969/j.issn.1673-8225.2012.10.020 BACKGROUND: Because of lacking of islet sources, the islet cells transplantation for the treatment of diabetes can not meet the clinical demand, so the differentiation of pancreas stem cells into islets in vitro has become a focus of the research. OBJECTIVE: To in vitro induce the mice pancreas stem cells into the islet-like cell clusters and to perform the relate measurement; to investigate the techniques and methods to differentiate the pancreas stem cells into the islets as well as the detection method.   METHODS: The mice pancreas stem cells were obtained in vitro, and then the joint inducer was used to induce the pancreas stem cells to differentiate into islets. The islet-like cell clusters were preformed with morphological observation, dithizone dyeing, RT-PCR and Western blot detection. RESULTS AND CONCLUSION: Through the observation of cell morphology and cell growth characteristics and immunocytochemistry staining, we obtained mice pancreas stem cells in vitro. The joint inducer was used to induce the mice pancreas stem cells to differentiate into islet-like cells, the cells were spherical with a more slender pedicle connected with the bottom, and the cells was stained iron red by dithizone dyeing. The results of RT-PCR and Western blot determined the expression of insulin mRNA and insulin protein of islet-like cells respectively. It confirms that the mice pancreas stem cells can be induced in vitro to differentiate into beta containing islet-like cell clusters. Related Articles | Metrics

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Small interfering RNA-mediated RhoA silencing optimizes the culture of fetal liver stem cells  Zhao Ge, Liu Wei-hui, Wang Tao, Wang Xing, Xia Ning, Yang Ping, Kou Ming-wen, Zhang Ning, Tao Kai-shan 2012, 16 (10):  1803-1807.  doi: 10.3969/j.issn.1673-8225.2012.10.021 Abstract ( 299 )   PDF (401KB) ( 298 )   Save BACKGROUND: Cell transplantation for disease treatment is one of interesting topics. To find a better source of seed cells and a more efficient amplification method is the basis for clinical application. OBJECTIVE: To determine whether the RhoA gene silencing in fetal liver stem cells can optimize liver stem cell cultures. METHODS: Fetal liver stem cells were in vitro cultured and were transfected by small interfering RNA (siRNA) to knock down the expression of RhoA. Then the cells were randomly divided into three groups. In the group A, fetal liver stem cells were non-treated. In the group B, fetal liver stem cells were transfected with randomly sequenced siRNA. In the group C, fetal liver stem cells were transfected with siRNA. RhoA gene and protein expression prior to and after transfection was detected by RT-PCR and western blot methods, respectively. Cellular proliferation was determined by MTT assay. The cell cycle was analyzed by flow cytometry. RESULTS AND CONCLUSION: Compared with groups A, B, the expression of RhoA gene and protein in the group C was significantly decreased (P < 0.05), the cell growth speed was significantly increased, cells in the G0/G1 phase of the cell cycle were decreased, and cells in the S phase were increased (P < 0.05). These findings suggest that the RhoA gene silencing can promote the proliferation of fetal liver stem cells and optimize the culture of fetal liver stem cells.  Related Articles | Metrics

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Effects of Dlx2 overexpression on cell cycle regulation and apoptosis of preosteoblast cells Sun Hao, Wang Xu-dong, Dai Jie-wen, Lu Jing-ting, Shen Guo-fang 2012, 16 (10):  1808-1812.  doi: 10.3969/j.issn.1673-8225.2012.10.022 Abstract ( 257 )   PDF (1157KB) ( 772 )   Save BACKGROUND: Dlx2 plays an important role in the regulation of cranial neural crest cells migration and craniofacial skeleton development. However, the effects of Dlx2 on apoptosis and cell cycle in osteoblast differentiation are not reported. OBJECTIVE: To investigate the effects of Dlx2 overexpression on cell cycle regulation and apoptosis in MC3T3-E1 preosteoblast cells differentiation. METHODS: Anti-retroviral pMSCV-puro-Dlx2 was transfected into MC3T3-E1 cells which were cultured in mineralization induced fluid. MC3T3-E1-Dlx2 cell lines stably overexpressing Dlx2 were constructed. Cell lines of Dlx2 over-expression were determined by RT-PCR and western blot. Apoptosis was detected by flow cytometry after Annexin V/PI and PI/RNase staining and cell cycle changes were detected by flow cytometry after PI/RNase staining. RESULTS AND CONCLUSION: MC3T3-E1-Dlx2 cell lines was successfully constructed. Cell cycle was blocked in G1/G0 phase (P < 0.05) and apoptosis was up-regulated (P < 0.05) in Dlx2 overexpression cell model. Dlx2 overexpression reduces cell proliferation and promotes cell differentiation to exercise functions. Related Articles | Metrics

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Application of acetaldehyde dehydrogenase in detection of endometrial stem cells Zhang Xiao-lu, He Yuan-li, Ma Ying 2012, 16 (10):  1813-1816.  doi: 10.3969/j.issn.1673-8225.2012.10.023 Abstract ( 283 )   PDF (309KB) ( 520 )   Save BACKGROUND: Stem-like cells have been found in human endometrium but the specificity marker has not been found. This kind of cells is difficult to be separated. OBJECTIVE: To investigate the usefulness of aldehyde dehydrogenase (ALDH) activity as a marker for endometrial stem cells. METHODS: Human endometrial cells were isolated and dissociated mechanically and enzymatically from human endometrium. Stromal and epithelial cells were separated by two series of filters. The ALDHhigh cells and ALDHlow cells were selected by flow cytometry based on ALDH activity. Limiting dilution culture was performed to culture these cells. Cell morphology and growth characteristics were observed. Flow cytometry analysis was used to identify the expression of c-kit, CD90, CD73 and CD29 markers in cultured cells. RESULTS AND CONCLUSION: The proportion of ALDHhigh cells was (1.88±0.17)% in stromal cells. After primary cultured for 15 days, the colony forming rate of the ALDHhigh cells reached 4.50%±0.82% which were significantly higher than ALDHlow cells (1.06±0.34)% (P < 0.05). Meanwhile, the large colony forming number of ALDHhigh cells was higher than that of ALDHlow cells (P < 0.05). The ALDHhigh cells and colony forming were not detected in epithelial cells. The expressions of c-kit, CD29, CD90 and CD73 cells were positive in endometrium colony forming cells. Results have proved that ALDH activity detection can select the endometrial stem cells. Related Articles | Metrics

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Human bone morphogenetic protein-2 recombinant adenovirus expression vector transfection of rabbit bone marrow mesenchymal stem cells    He Chun-lei, Ji Guang-lin, Liu Wu-yang, Wu Dong-bao, He Cheng, Wang Mao-yuan, Ye Yong-jun, Mo Jian-wen, Gao Hui 2012, 16 (10):  1817-1821.  doi: 10.3969/j.issn.1673-8225.2012.10.024 Abstract ( 259 )   PDF (510KB) ( 279 )   Save BACKGROUND: The adenovirus vector structured by GatewayTM has made up the deficiencies of exogenous cytokine application.  OBJECTIVE: To observe the target gene expression and osteoblastic differentiation of human bone morphogenetic protein-2 (hBMP-2) recombinant adenovirus expression vector (Ad-BMP-2/GFP) after transfected with bone marrow-derived mesenchymal stem cells (rBMSCs). METHODS: To isolate and culture, purify rBMSCs through the density gradient centrifugation combined with adherent cultivation; the Ad-BMP-2/GFP adenovirus vector constructed by GatewayTM technology was transfected with rBMSCs. RESULTS AND CONCLUSION: Osteocalcin was the osteoblast-specific product which could be expressed on the transfected rBMSCs detected by RT-PCR method; hBMP-2 gene was positive at mRNA and protein levels. Rabbit bone marrow stromal cells were successfully separated and purified. Ad-BMP-2 can efficiently transfect rBMSCs and the BMP-2 can be expressed in BMSCs efficiently. Related Articles | Metrics

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Effect of 5-fluorouracil on the expression of Oct4 and Bmi-1 in human breast cancer cell line MCF7   Yan Li1, Shi Li-Gang1, Xu Zheng-Shun1, Li Hui-Xiang2 2012, 16 (10):  1822-1826.  doi: 10.3969/j.issn.1673-8225.2012.10.025 Abstract ( 333 )   PDF (454KB) ( 338 )   Save BACKGROUND: Several studies suggest that occurrence and development of breast cancer is a progressional process in which many factors participate, and breast cancer stem cells are thought to be the root cause that may initiate the growth of breast cancer recurrence and far metastasis after neoadjuvant chemotherapy. OBJECTIVE: To investigate the influence of 5-fluorouracil (5-Fu) on the expression of Oct4 and Bmi-1 in human breast cancer cell line MCF7. METHODS: MCF7 was treated with 0.1 mg/L 5-Fu, the expression of Oct4 and Bmi-1 in untreated cells and six generations of treated cells was detected with reverse transcription-polymerase chain reaction, and the expression of Oct4 in untreated cells and five generations of treated cells was detected by immunocytochemistry. RESULTS AND CONCLUSION: After 5-Fu treatment, Oct4 and Bmi-1 mRNA expression in the third and fifth generations of treated cells was significantly increased, and Oct4 protein during first to fifth generations of cells had a certain degree of regularity. These findings suggest that there may be cancer stem cell-like cells in MCF7 and 5-Fu might increase the number of cancer stem cell-like cells in MCF7. Related Articles | Metrics

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Herpes simplex virus thymidine kinase expressing mesenchymal stem cells in combination with Ganciclovir in targeted-gene therapy for nasopharyngeal carcinoma   Zhong Pin-neng, Wen Zhong, Shen Cong-xiang, Wang Hai-li, Wang Jun-qi 2012, 16 (10):  1827-1832.  doi: 10.3969/j.issn.1673-8225.2012.10.026 Abstract ( 381 )   PDF (655KB) ( 312 )   Save BACKGROUND: Recent studies have confirmed that mesenchymal stem cells (MSCs) can be used as targeted-gene delivery vehicles for cancer gene therapy. OBJECTIVE: To investigate the therapeutic effects of herpes simplex virus thymidine kinase expressing mesenchymal stem cells (TK- MSCs) in combination with Ganciclovir (GCV) on nasopharyngeal carcinoma. METHODS: The pGL3 -EGFP-TK plasmid was constructed and transfected into Sprague-Dawley rat MSCs by Lipofectamine™ 2000. Cell proliferation was determined by CCK-8 method. Tumor-homing of TK-MSCs was analyzed by Transwell inserts. BALB/C nude mice were inoculated with TK-MSCs to observe the tumorigenicity. Human nasopharyngeal carcinoma cells 5-8F were interfered with TK-MSCs/GCV to investigate the cell viability and the bystander effect. RESULTS AND CONCLUSION: TK gene was transfected into MSCs successfully. There was no significant difference in cell proliferation between TK-MSCs and MSCs (P > 0.05). TK-MSCs maintained their tumor-homing and had no tumorigenicity. TK-MSCs/GCV significantly inhibited the growth of 5-8F cells (P < 0.01). TK-MSCs/GCV suicide gene therapy system exhibits significant inhibitory effects on growth of 5-8F cells, suggesting that MSCs can be used as delivery vehicles for targete-gene  therapy of nasopharyngeal carcinoma. Related Articles | Metrics

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Transplantation of bone marrow mesenchymal stem cells improves cognitive function of rats with chronic cerebral ischemia and decreases Nogo-A and NgR protein expression in the hippocampus    Wang Yan-ling, Yan Yu, Song Bo, Zhang Hui-li, Gong Guang-ming, Chen Si, Fang Hui, Xu Yu-ming 2012, 16 (10):  1833-1836.  doi: 10.3969/j.issn.1673-8225.2012.10.027 Abstract ( 222 )   PDF (358KB) ( 358 )   Save BACKGROUND: Transplantation of bone marrow mesenchymal stem cells can decrease Nogo-A and NgR expression in rats with spinal cord injury and cerebral infarction. OBJECTIVE: To investigate the effects of transplantation of bone marrow mesenchymal stem cells on cognitive function of rats with chronic cerebral ischemia and Nogo-A and NgR protein expression in the hippocampus. METHODS: Thirty Sprague-Dawley rats were randomly divided into sham-surgery, model and bone marrow mesenchymal stem cells groups. Rats in the latter two groups were established into model of chronic cerebral ischemia by permanent ligation of bilateral common carotid artery. In the bone marrow mesenchymal stem cells group, at 7 days after chronic cerebral ischemia induction, bone marrow mesenchymal stem cells (1×107) isolated by ficoll density gradient centrifugation were transfused into rats via the tail vein. RESULTS AND CONCLUSION: Compared with sham-surgery group, the average escape latency was significantly shortened (P < 0.01), the number that rats passed through the platform was significantly increased (P < 0.05), and Nogo-A and NgR expression in the hippocampal tissue was significantly decreased (P < 0.05) in the bone marrow mesenchymal stem cells group. These findings suggest that transplantation of bone marrow mesenchymal stem cells protects spatial learning and memory abilities of rats with chronic cerebral ischemia. This effect is possibly related to decreased Nogo-A and NgR protein expression. Related Articles | Metrics

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Transplantation of human umbilical cord-derived mesenchymal stem cells improves hepatic fibrosis in rats with carbon etrachloride-induced hepatic cirrhosis Liu Ying1, Shi Zhan-li2, Zhao Zong-ze2, Guo Sheng-nan1, Xu Jin-kai1, Li Dong-jie 2 2012, 16 (10):  1837-1840.  doi: 10.3969/j.issn.1673-8225.2012.10.028 Abstract ( 298 )   PDF (375KB) ( 432 )   Save BACKGROUND: The feasibility and efficacy of stem cell transplantation as a new method in treatment of hepatic cirrhosis have been rarely reported at home and abroad. OBJECTIVE: To investigate the effects of human umbilical cord-derived mesenchymal stem cells (hUCMSCs) on hepatic fibrosis in rats with carbon etrachloride-induced hepatic cirrhosis. METHODS: Thirty-two Wistar rats were randomly divided into two groups. Rats in the control group were injected with normal saline via the tail vein. Rats in the hepatic cirrhosis group were established into rat models of hepatic cirrhosis by cutaneous injection of carbon etrachloride peanut oil for 8 weeks. The hepatic cirrhosis rats were then randomly sub-divided into three groups: 8-week hepatic cirrhosis, normal saline, hUCMSCs transplantation. Rats in the normal saline and hUCMSCs transplantation subgroups were administered normal saline and hUCMSCs suspension respectively via the tail vein. Rats in the 8-week hepatic cirrhosis received no intervention. RESULTS AND CONCLUSION: Serum alkaline phosphatase level was significantly lower in the control group than in the other three subgroups (P < 0.05). Serum albumin and cholesterol levels in the hUCMSCs group were similar to those of control group (P > 0.05), but they were significantly higher compared with 8-week hepatic cirrhosis and normal saline group (P < 0.05). In the 8-week hepatic cirrhosis group, a large amount of hyperplastic collagenous fiber was presented in the liver tissue and the frame of the pseudolobuli formed. The normal saline group and 8-week hepatic cirrhosis group exhibited similar histological changes. Scattered green antinuclear antibody-positive cells in the liver were only observed in the hUCMSCs group. The amount of hyperplastic collagenous fiber was significantly lower in the hUCMSCs transplantation group than in the normal saline group. These findings suggest that transplantation of human umbilical cord-derived mesenchymal stem cells via the tail vein can obviously improve hepatic fibrosis in rats with carbon etrachloride-induced hepatic cirrhosis. Related Articles | Metrics

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Treatment of myocardial infarction by transplantation of bone marrow mesenchymal stem cells transfected by adenovirus-mediated vascular endothelial growth factor and hepatocyte growth factor   Chen Guo-xiang1, Hua Ping2, Xiong Li-hua2, Chen Ju2, Pan De-qiang1 2012, 16 (10):  1841-1845.  doi: 10.3969/j.issn.1673-8225.2012.10.029 Abstract ( 216 )   PDF (442KB) ( 274 )   Save BACKGROUND: The bone marrow mesenchymal stem cells (BMSCs) can differentiate into myocardial cells and promote angiogenesis. However, the cell factor excreted by BMSCs in the early period of transplantation could not promote the differentiation and regeneration. OBJECTIVE: To investigate the effects of BMSCs transplantation transfected by adenovirous mediated vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) gene therapy on the tissue repair and angiogenesis in rabbits with myocardial infarction． METHODS: Rabbit BMSCs were isolated, cultured and purified in vitro, then labeled with BrdU and transfected by adenovirus mediated VEGF and HGF gene. 50 rabbits were used to establish the acute myocardial infarction models. Four weeks later, 50 rabbit models were randomly divided into five groups: group A was injected with BMSCs/Ad.VEGF+HGF; group B was injected with BMSCs/Ad.HGF; group C was injected with BMSCs/Ad.VEGF; group D was injected with BMSCs; group E was control group, it was injected with the same concentration of serum-free IMDM medium. The differentiation and angiogenesis of BMSCs were observed at 4 weeks after transplantation. Doppler echocardiography was performed to detect the changes on heart function． RESULTS AND CONCLUSION: Except for control group, the heart function was obviously improved after transplantation in group A, B, C and D (P < 0.05), and the improvement was the best in A group. Some BrdU stained positive BMSCs could differentiate into the endothelial cells and incorporate into the coronary capillaries in the infracted region. Compared to the control group, the other four groups could facilitate the angiogenesis (P < 0.05), especially in the group A. BMSCs implantation combining with VEGF and HGF gene therapy can obviously promote the myocardial regeneration angiogenesis, and can improve the heart function. Related Articles | Metrics

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Effects of ganglioside combined with neural stem cells transplantation on hippocampal neuron following brain injury in rats  Li Hong-xing1, Zhao Zong-mao2, Jin Yao-dong3, Wang Jing4, Yuan Guo-fu5 2012, 16 (10):  1846-1850.  doi: 10.3969/j.issn.1673-8225.2012.10.030 Abstract ( 229 )   PDF (425KB) ( 353 )   Save BACKGROUND: The research about the application of neural stem cells (NSCs) transplantation on the treatment of neurological dysfunction after brain injury is a hot spot. OBJECTIVE: To investigate the effect of ganglioside combined with NSCs transplantation on hippocanipal neuron following traumatic brain injury (TBI) in rats． METHODS: Sixty-six healthy Wistar rats were selected to establish the hippocanipal neuron model cause by TBI using improved Feeney method. After injured for 24 hours, the surviving 60 rats were randomly divided into 3 groups, the TBI group was injected with 1 mL DMEM/F12 medium; the NSCs group was injected with 1×1010/L NSCs suspension; the NSCs+ganglioside group was injected with 1×1010/L NSCs suspension and then intraperitoneal injection of 30 mg/kg ganglioside aqueous solution, once per day and lasted for 3 days.  RESULTS AND CONCLUSION: Fluorescence microscope observation showed that the NSCs labeled by PKH26 were spherical and presented with red and uniform-distributed fluorescence. Flow cytometric detection revealed that the positive percentage of PKH26 labeled NSCs was over 95%. The average time of escape latency was gradually decreased in each group. However, from the 3rd to 5th day, time in NSCs+ganglioside group was shorter than that in NSCs transplantation group (P < 0.05) and compared with TBI group, time in NSCs+ganglioside group was decreased significantly (P < 0.01). In addition, the frequency of platform passing and the percentage of swimming distance in target quadrant and the total distance were highest in NSCs+ganglioside group (P < 0.05). The number of PKH26-positive cells and neurons in the section stained by hematoxylin-eosin as well as the mRNA expression of brain-derived neurotrophic factor in NSCs+ganglioside group was more than that in NSCs group, and those factors in NSCs group were more than those in TBI group (P < 0.05). NSCs transplantation plays a role in protecting against hippocampal neuronal death following TBI, with the affiliation of ganglioside claim synergism effects.   Related Articles | Metrics

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Brain-derived neurotrophic factor expression in the spinal dorsal horn and dorsal root ganglion in a rat model of sciatic nerve injury after intrathecal transplantation of neural stem cells  Zhang Min-hao1, Hu Yi-min1, Zhang Hong2, Hu Yu-ping1, Liu Qing-zhen1, Wang Hong-chao2, Li Wei-yan1 2012, 16 (10):  1851-1855.  doi: 10.3969/j.issn.1673-8225.2012.10.031 Abstract ( 269 )   PDF (399KB) ( 297 )   Save BACKGROUND: The model of sciatic nerve injury can be used to test the hyperalgesia and paraesthesia caused by nocuous heat stimulation and mechanical stimulation. OBJECTIVE: To investigate the expression of brain-derived neurotrophic factor (BDNF) in the spinal dorsal horn and dorsal root ganglion in a rat model of sciatic nerve injury after intrathecal transplantation of neural stem cells. METHODS: Seventy-two adult Sprague-Dawley rats were randomly divided into sham-operation, control and experimental groups. Rat model of sciatic nerve injury was established in the control and experimental groups. In the sham-operation group, only sciatic nerve was exposed, without ligation. Intratheal transplantation was performed at 3 and 10 days after induction of sciatic nerve injury. 30 μL of neural stem cell suspension was injected into the experimental group, and 30 μL cell culture solution was injected into the sham-operation and control groups.   RESULTS AND CONCLUSION: Compared with sham-operation group, mechanical withdrawal threshold and thermal withdrawal latency were gradually reduced at 3 days after transplantation, decreased to the lowest level at 7 days (P < 0.01) and recovered to pre-transplantation level at 21 days in the control and experimental groups. At 7 and 14 days after transplantation, mechanical withdrawal threshold and thermal withdrawal latency were significantly increased in the experimental group than in the control group (P < 0.01). Compared with control group, at 7, 14 and 21 days after transplantation, BDNF expression was significantly decreased in the experimental group (P < 0.05). At 14 and 21 days after transplantation, BDNF expression was significantly higher in the experimental group than in the control group (P < 0.05). These findings show that intrathecal transplantation of neural stem cells can prevent and treat the neuropathic pain from peripheral nerve injury by increasing BDNF expression in the spinal dorsal horn and dorsal root ganglion. Related Articles | Metrics

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Bcl-2 and Bax expression in mice with collagen-induced arthritis following heterogenic umbilical cord blood stem cell transplantation*☆ Niu Guang-hua, Gao Yu-jie, Sun Xu, Du Jing, Ming Cai-rong, Gao Ming-li 2012, 16 (10):  1856-1860.  doi: 10.3969/j.issn.1673-8225.2012.10.032 Abstract ( 344 )   PDF (400KB) ( 301 )   Save BACKGROUND: The occurrence and development of rheumatoid arthritis were strongly associated with unbalance proliferation and apoptosis of synovial cells and lymphocytes. Some synovial cell apoptosis was abnormal. OBJECTIVE: To observe effects of heterogenic double umbilical cord blood stem cell transplantation on Bcl-2 and Bax expression in mice with type Ⅱ collagen-induced arthritis. METHODS: C57BL/6(H-2b) mice were induced with Freund’s complete adjuvant and type Ⅱ collagen to establish mouse models of type Ⅱ collagen-induced arthritis. At 2 days after secondary immunity incubation, mice were injected with saline via tail vein in the model and normal control groups. Umbilical cord blood hemopoietic stem cells were injected into mouse tail vein in the UBSCs transplantation groups (single dose: 2×106/50 g; double dose: 1×106/50 g each, totally 2×106/50 g). Mice were intragastrically administrated methotrexate in the methotrexate positive control group, 0.017 5 g/kg once, once every 5 days, totally six times. RESULTS AND CONCLUSION: Joint tissue below knee and elbow was obtained at 42 days following transplantation. Histopathology displayed that smooth and glossy articular surface, no inflammatory cell infiltration in the synovial layer and normal chondrocytes in the normal control group. Hyperplasia, a lot of inflammatory cell infiltration and damaged cartilage surface were visible in the model group. Slight hyperplasia and a little inflammatory cell infiltration were detectable in the methotrexate positive control group and single UBSCs transplantation group. Double UBSCs transplantation group exhibited smooth and glossy articular cartilage surface, no damage, a little inflammatory cell infiltration. Immunohistochemistry demonstrated that Bcl-2 and Bax expression was lower in the double UBSCs transplantation group compared with single UBSCs transplantation group (P < 0.05). Compared with normal control group, no significant difference was detected (P > 0.05). Results suggest that double umbilical cord blood stem cells can induce synovial cell apoptosis in mice with type Ⅱ collagen-induced arthritis in a certain number and action time, and protect synovial membrane against damage. Related Articles | Metrics

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Bone marrow mesenchymal stem cells differentiation into cardiomyocyte-like cells induced by 5-azacytidine and astragaloside Ⅳ***☆ Xian Shao-xiang, Yang Zhong-qi, Qin Jia-jia, Huang Xi-wen, Sun Jing-he 2012, 16 (10):  1861-1865.  doi: 10.3969/j.issn.1673-8225.2012.10.033 Abstract ( 362 )   PDF (416KB) ( 374 )   Save BACKGROUND: 5-azacytidine (5-Aza) has been frequently used to induce bone marrow mesenchymal stem cells (BMSCs) differentiation into cardiomyocyte. OBJECTIVE: To observe expression of cardiomyocyte-related receptors in cardiomyogenic differentiation of rat BMSCs. METHODS: BMSCs of passage three were assigned to four groups: group Ⅰ: L-DMEM solution alone was replaced; Ⅱ: L-DMEM solution was replaced after induction of 100 mg/L AST+5 µmol/L 5-Aza for 24 hours; group Ⅲ: L-DMEM solution was replaced after induction of 10 µmol/L 5-Aza for 24 hours; and group Ⅳ: L-DMEM solution was replaced after induction of 5 µmol/L 5-Aza for 24 hours. Culture medium was replaced every 3 days in each group. Differentiated cells were identified after 30 days of induction. RESULTS AND CONCLUSION: Expression of cardiomyocyte specific proteins Nkx2.5, cTnT and Desmin was detected in groups Ⅲ, Ⅳ and Ⅱ after induction compared with group Ⅰ , with significant differences (P < 0.01). The amount of cTnT and Desmin expression expression was significantly higher in groups Ⅱ and Ⅲ compared with group Ⅳ (P < 0.01). The level of Nkx2.5 expression was significantly higher in groups Ⅱ (P < 0.01) and Ⅲ (P < 0.05) compared with group Ⅳ. No Nkx2.5, cTnT and Desmin espression was detected in group Ⅰ. After induction for 2 weeks, cells with spontaneous contractility were observed in groups Ⅱ and Ⅲ, indicating differentiation towards cardiomyocyte after induction. Results demonstrated that induction effects were similar between 100 mg/L AST+5 µmol/L 5-Aza and 10 µmol/L 5-Aza. This may contribute to cytoprotective effects of AST, which can promote vascular endothelial cell proliferation, enhance celss tolerance to 5-Aza-induced cytotoxicity and upregulate cardiac-specific protein expression. Related Articles | Metrics

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Diverse expression patterns of microRNA-155 between bone marrow-derived M1 and M2 macrophages   Liang Yan-bing, Wang Zhong-hua, Tang Hao, Ma Zhong-fu 2012, 16 (10):  1866-1870.  doi: 10.3969/j.issn.1673-8225.2012.10.034 Abstract ( 252 )   PDF (392KB) ( 439 )   Save BACKGROUND: Previous studies of microRNA-155 expression and function in macrophage predominantly focus on total mononuclear macrophage. OBJCTIVE: To evaluate the expression difference of microRNA-155 in M1 and M2 macrophages. METHODS: Bone marrow-derived macrophages were induced to differentiate towards M1 and M2 macrophages using interferon-γ (100 U/mL) + lipopolysaccharide (5 ng/mL) and interleukin-4 (10 ng/mL), respectively. RESULTS AND CONCLUSION: The differentiation proportion of M1 and M2 macrophages detected by flow cytometry was 91% and 95%, respectively. Reverse transcription PCR method showed that inducible nitric oxide synthase (iNOS) mRNA was highly expressed in M1 macrophages, but it was hardly expressed in M2 macrophages. Arginase-1 and found in inflammatory zone 1 mRNA expression was highly expressed in M2 macrophages, but the expression was low in M1 macrophages. Tumor necrosis factor α mRNA expression was significantly higher in M1 macrophages than in M2 macrophages, on the contrary, interleukin 10 mRNA expression was significantly higher in M2 macrophages than in M1 macrophages (P < 0.05). Real-time fluorescence quantitative PCR showed that microRNA-155 expression in M1 macrophages was significantly higher compared with that in M2 macrophages (P < 0.05). These findings suggest that MiRNA-155 can serve as a useful maker for differential diagnosis of M1 and M2 macrophages. Related Articles | Metrics

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Research progress in mesenchymal stem cells transplantation for treatment of multiple sclerosis Liu Ying, Hou Zong-liu 2012, 16 (10):  1869-1892.  doi: 10.3969/j.issn.1673-8225.2012.10.039 Abstract ( 333 )   PDF (410KB) ( 319 )   Save BACKGROUND: Multiple sclerosis (MS) is a neurodegenerative disease of the central nervous system and has a complex etiology that is not fully understood. The clinical course of MS is heterogeneous, and the traditional therapeutic interventions for MS have not been convincingly shown to change the natural course of the disease. OBJECTIVE: To summarize the mechanism and application of mesenchymal stem cells transplantation for the treatment of multiple sclerosis and raise some questions that should be settled. METHODS: Kjmed databases were searched by the first author using key words of “mesenchymal stem cells, encephalomyelitis, autoimmune, experimental, multiple sclerosis” in English from 2000 to 2011. Literatures that related with the mesenchymal stem cells for the treatment of MS were included, and the repetitive researches were excluded. RESULTS AND CONCLUSION: A total of 104 documents were retrieved, and 33 articles were retained after deleting unrelated and repetitive ones. To date, the research had showed that mesenchymal stem cells were multipotent stem cell with the characteristics of immune regulatory and neuroprotective properties, and had showed the initial therapeutic effect on the treatment of MS. Related Articles | Metrics

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Effect of shear stress on the copper-zinc superoxide dismutase gene expression and activity in human endothelial progenitor cells  Yang Zhen1, Lai Guang-hua1, Xia Wen-hao1, Luo Chu-fan2, Wang Jie-mei1, Chen Long1, Liao Xin-xue1, Jin Ya-fei1, Tao Jun1 2012, 16 (10):  1871-1874.  doi: 10.3969/j.issn.1673-8225.2012.10.035 Abstract ( 275 )   PDF (386KB) ( 367 )   Save BACKGROUND: Shear stress is an important non-pharmacological method to regulated endothelial progenitor cells (EPCs). However, the effect of shear stress on the anti-oxidative activity of EPCs is not clear. OBJECTIVE: To observe the effect of shear stress on the copper-zinc superoxide dismutase (Cu/Zn-SOD) gene expression and activity in EPCs, in order to investigate the regulatory effects of shear stress on the EPCs function. METHODS: The peripheral blood mononuclear cells of healthy adult were inducted into EPCs. Then, they were divided into four different experimental groups which included stationary group, low-flow shear stress group (0.05 mN /cm2), medium-flow shear stress group (0.15 mN/cm2) and high-flow shear stress group (0.25 mN/cm2). RESULTS AND CONCLUSION: The peripheral blood mononuclear cells were differentiated into EPCs. They presented typical “spindle-shaped” appearance, and were positively labeled by acetylated-LDL, lectin, FLK-1 and VWF. The Cu/Zn-SOD activity of human EPCs in shear stress treatment groups was higher than that in stationary group. The higher the shear stress, the higher the Cu/Zn-SOD activity of human EPCs. Quantitative real-time PCR indicated that shear stress could increase the CuZn-SOD mRNA expression of EPCs, the higher the shear stress, the stronger the CuZn-SOD mRNA expression of EPCs. In vitro shear stress could upregulate the gene expression of Cu/Zn-SOD in EPCs, increase the Cu/Zn-SOD activity in human EPCs, which contributes to enhance the anti-oxidative activity of EPCs. Therefore, enhanced shear stress within the physiological range could regulate the function of EPCs, which can act as an important strategy of enhancing their therapeutic approach for vascular endothelial injury. Related Articles | Metrics

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Immunofluorescence for dynamic monitoring of PML-RARα fusion protein in patients with acute promyelocytic leukemia Qiu Fei, Zhang Hui, Zhao Xu-jie, Zhu Xue-hua, Wang Kan-kan, Zhang Ji 2012, 16 (10):  1875-1878.  doi: 10.3969/j.issn.1673-8225.2012.10.036 Abstract ( 318 )   PDF (519KB) ( 359 )   Save BACKGROUND: Real-time quantitative RT-PCR can directly reflect the expression change of fusion gene, but it cannot directly reflect the relative expression of intracellular PML-RARα fusion protein in patients with acute promyelocytic leukemia (APL) before and after all-trans retinoic acid treatment. OBJECTIVE: To investigate the reliability of immunofluorescence technique in judging the early-stage therapeutic effects of all-trans retinoic acid treatment in APL by dynamic monitoring PML-RARα fusion protein. METHODS: Immunofluorescence technique was used for detection of PML-RARα protein expression in typical APL cell strain BN4 cells or NB4 cells of patients with APL following all-trans retinoic acid treatment. Real-time quantitative RT-PCR was used for detection of intracellular PML-RARα mRNA expression as control to investigate the reliability of immunofluorescence technique. RESULTS AND CONCLUSION: Immunofluorescence staining results showed that before all-trans retinoic acid treatment, a large amount of red fusion protein PML-RARα of different size and irregular morphology was observed in BN4 cells in vitro cultured or from patients’ bone marrow; after all-trans retinoic acid treatment, PML-RARα fusion protein in the NB4 cells gradually became regular and was gradually scattered with prolongation of treatment time. Real-time quantitative RT-PCR results showed that the change tendency of PML-RARα mRNA expression was in accordance with immunofluorescence results. This suggests that immunofluorescence for detection of PML-RARα expression can be used for directly judging the therapeutic effects of all-trans retinoic acid treatment.  Related Articles | Metrics

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Purification of human peripheral blood monocytes on gelatin-coated surface and stimulation into dendritic cells  Ma Ming-ying1,2, Jin Zhi-ping1, Wang Wei1, Guo Zhi1, Wu Bin3,4, Tan Xiao-hua1,4 2012, 16 (10):  1879-1883.  doi: 10.3969/j.issn.1673-8225.2012.10.037 Abstract ( 375 )   PDF (315KB) ( 278 )   Save BACKGROUND: There have been no standard procedures regarding how to simply and efficiently separate and purify human peripheral blood monocytes and stimulate them into dendritic cells. OBJECTIVE: To investigate the efficacy of purification of human peripheral blood monocytes on gelatin-coated surfaces, analyze the phenotype of dendritic cells generated by these monocytes, and make a comparison with conventional plastic adhesion method. METHODS: Human peripheral blood mononuclear cells were harvested by Ficoll-Hypaque gradient centrifugation. Gelatin group and common plastic group were designated according to coating flasks with or without gelatin. Monocytes were harvested from each group and then were stimulated into dendritic cells. The number of monoctyes, CD14 positive rate of monocytes, contaminated T, B lymphocytes, the expression of CD1a, CD83 on immature and mature dendritic cells were determined. Cell viability was determined by trypan blue staining. Platelet contamination was compared between gelatin and common plastic groups. RESULTS AND CONCLUSION: The number of monocytes and CD14 positive rate were significantly higher in the gelatin group than in the common plastic group (P < 0.05). Contaminated T, B lymphocytes were significantly more in the common plastic group than in the gelatin group (P < 0.05). There were no significant differences in cell viability and dendritic cell phenotype between gelatin group and common plastic group (P > 0.05). Platelet contamination rate was significantly lower in the gelatin group than in the common plastic group. Gelatin-coated surfaces can effectively and simply isolate monocytes and successfully stimulate them into mature dendritic cells. Related Articles | Metrics

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Tanshinone-ⅡA induced differentiation of rat bone marrow mesenchymal stem cells into neuron-like cells in vitro after pre-induced by basic fibroblast growth factors Li Yu-mei1, Yu De-li2, Liu Lai-bing3, Yu Zi-jiang1 2012, 16 (10):  1884-1888.  doi: 10.3969/j.issn.1673-8225.2012.10.038 Abstract ( 270 )   PDF (526KB) ( 371 )   Save BACKGROUND: Whether bone marrow mesenchymal stem cells (BMSCs) induced by traditional Chinese medicine could differentiate into neuron-like cells remains poorly understood.                              OBJECTIVE: To identify the efficiency of tanshinone-ⅡA containing traditional Chinese medicine on the differentiation of rat BMSCs into neuron-like cells in vitro after pre-induced by basic fibroblast growth factors.                                             METHODS: One month old healthy male Wistar rats were used in this experiment, and BMSCs were separated and purified by Percoll medium density gradient centrifugation, repeated subculture and differential adherence methods. Passage 3 BMSCs were differentiated into neuron-like cells induced by basic fibroblast growth factors (bFGF) and tanshinone-ⅡA. All-trans retinoic acid serum-free medium induced cells and non-induced cells served as controls.                                             RESULTS AND CONCLUSION: ①BMSCs could stably express the stem cells specific markers CD44, and the positive rate was (91.00±1.58)%. And the negative expressions of CD34 and CD45 were observed in the BMSCs by immunohistochemistry and flow cytomtry and the purity of stem cells was up to 95.5%. ② After directional differentiation and cultivation, typical neuron-like cells could be seen under scanning electron microscope. Positive expression of NSE was detected by immunohistochemistry and the possitive rate was (75.6±2.31)%. Nestin expression was gradually decreased to negative as well as the expression of glial fibrillary acidic protein. The results suggested that tanshinone-ⅡA can directly and efficiently induce BMSCs differentiating into neuron-like cells after pre-induced by basic fibroblast growth factors.   Related Articles | Metrics

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Derivation of Schwann cells from stem cells Derivation of Schwann cells from stem cells 2012, 16 (10):  1893-1896.  doi: 10.3969/j.issn.1673-8225.2012.10.040 Abstract ( 265 )   PDF (386KB) ( 395 )   Save BACKGROUND: Schwann cells play an important role in the recovery and regeneration of peripheral nervous system (PNS) injury. However, Schwann cells are not easy to obtain directly which lead the limitation on clinical application.  OBJECTIVE: To review the effect of Schwann cells on nervous system injury and regeneration process, and the potency of other stem cells to differentiate into Schwann cells. METHODS: A computer-based research on the Pubmed database was conducted to search the articles on Schwann cells participating in the regeneration of nervous system injury and the identification and application of Schwann cells differentiated from other stem cells, the key words were “Schwann cells, nervous system, stem cells”. The repetitive articles were eliminated; each articles and quotations were reviewed during the process of preliminary examination. A total of 29 articles were included according to the inclusion criteria. RESULTS AND CONCLUSION: Schwann cells provide a suitable microenvironment for repair and regeneration through the secretion of various cytokines and trophic factors, thereby promote the axonal regeneration. Meanwhile, they can grow around the newborn axons and form the myelin through the mechanism of proliferation and differentiation. A large number of Schwann cells are required to repair the nervous system, but Schwann cells are not easy to directly obtain. Variety of stem cells, including embryonic stem cells, mesenchymal stem cells, neural crest stem cells and olfactory ensheathing cells can be induced to differentiate into Schwann cell-like cells in certain conditions which provide new hope for the treatment of nervous system injury. Among them, the researches on the mesenchymal stem cells attract the most attention.  Related Articles | Metrics

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Autologous hematopoietic stem cell transplantation in combination with allogenetic cytokine induced killer cells and interleukin-2 infusion for treatment of high risk acute myelocytic leukemia in two cases  Tu San-fang, Song Chao-yang, Li Yu-hua, Guo Kun-yuan, Huang Yu-xian 2012, 16 (10):  1897-1900.  doi: 10.3969/j.issn.1673-8225.2012.10.041 Abstract ( 360 )   PDF (337KB) ( 366 )   Save BACKGROUND: The relapse rate of patients with high risk acute myelocytic leukemia is very high after treated with autologous hematopoietic stem cells transplantation (AHSCT). How to decrease the relapse rate is still difficult. OBJECTIVE: To investigate the clinical efficacy of AHSCT combined with allogenentic cytokine-induced killer cells (CIK cells) and interleukin-2 (IL-2) infusion for treatment of high risk acute myelocytic leukemia (AML). METHODS: Two patients with high risk AML were treated with AHSCT at complete remission first time (CR1) after induction and consolidation chemotherapy. One month after AHSCT, two patients were infused with allogenetic CIK cells for 3-4 courses, with one course for half a year, 5 times per course and once per day. Subcutaneous injection with IL-2 was finished at half an hour before infusing CIK cells every time, the subcutaneous injection of IL-2 was preformed once every 2 days after the first infusion and once every 3 days after half a year, once every 4 days after one year, and once a week after one and half a year. The injection was maintained for half a year to prevent the relapse of AML. RESULTS AND CONCLUSION: After the two patients were treated with allogenentic CIK cells infusion and subcutaneous injection of IL-2 after AHSCT, there were no side effects appeared such as fever, shivering and erythra, at the same time there were no bone marrow depression and graft versus host disease appeared. The sustained remission time of the two patients was 20 months and 2 years respectively without relapse after AHSCT until now. The first time we find that the patients with high risk AML can acquire disease-free survival longtime with the treatment of AHSCT in combination with immunotherapy of CIK cells and IL-2 when they have no suitable donors. Related Articles | Metrics