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    05 November 2015, Volume 19 Issue 45 Previous Issue    Next Issue
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    Effect of adipose-derived stem cells modified by insulin-like growth factor I gene on TWEAK/Fn14 signaling pathway
    Liu Yu-ping, Wang Ming-ming, Liu Tao, Li Ming, Yu Guang-rong
    2015, 19 (45):  7217-7223.  doi: 10.3969/j.issn.2095-4344.2015.45.001
    Abstract ( 126 )   PDF (1008KB) ( 261 )   Save

    BACKGROUND: Because insulin-like growth factor I has the ability to induce mesenchymal stem cells into chondrocytes, we hypothesized that the chondrogenic differentiation of adipose-derived stem cells can be improved via insulin-like growth factor I transfection.

    OBJECTIVE: To investigate the influence of insulin-like growth factor I transfection on chondrogenic potential of adipose-derived stem cells in vitro and tumor necrosis factor-like weak inducer of apoptosis (TWEAK)/fibroblast growth factor-inducible 14 (Fn14) signaling pathway.
    METHODS: Recombinant lentivirus plasmid pLVX-IGF-I-IRES-ZsGreenl was constructed and transferred into passage 3 adipose-derived stem cells, and then these stem cells were induced to differentiate into chondrocytes (experimental group 1). Meanwhile, the cells transfected with pLVX-IRES-ZsGreenl were taken as green fluorescent protein/adipose mesenchymal stem cell group (experimental group 2), and those with no transfection acted as control group.
    RESULTS AND CONCLUSION: The mRNA expression of TWEAK was reduced in the experimental group 1 as compared with the other two groups, but the mRNA expressions of insulin-like growth factor I, Col2a1 and Sox9 were up-regulated in the experimental group 1. At the same time, the protein expression of matrix metalloproteinase-3 and TWEAK were down-regulated, while the protein expression of Col2a1 was increased in the insulin-like growth factor I-transfected cells in contrast to the cells modified with pLVX-IRES-ZsGreenl or with no transfection. These findings indicate that pLVX-IGF-I-IRES-ZsGreenl transfection of adipose-derived stem cells results in a higher expression of insulin-like growth factor I, and down-regulates the expression of TWEAK mRNA and protein, which improves the differentiation of adipose-derived mesenchymal stem cells into chondrocytes. 
    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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    Effect of silk fibroin/hydroxyapatite scaffold on the viability and osteogenic properties of adipose-derived stem cells under osteogenic induction
    Liu Hao, Chu Ya-wei, Ding Tao, Cheng Li, Zhu Hao-ming
    2015, 19 (45):  7224-7229.  doi: 10.3969/j.issn.2095-4344.2015.45.002
    Abstract ( 120 )   PDF (923KB) ( 307 )   Save

    BACKGROUND: Adipose-derived stem cells under osteogenic induction can be combined with biodegradable silk fibroin/hydroxyapatite scaffold, which is expected to develop a new biocompatible and osteogenic bone fusion material.

    OBJECTIVE: To study the effect of silk fibroin/hydroxyapatite composite on the viability and osteogenic properties of adipose-derived stem cells after osteogenic induction.
    METHODS: Adipose-derived stem cells were obtained from rat’s fat tissue, then adherently cultured, proliferated and passaged in vitro. Passage 3 cells were cultured in conditioned medium for osteogenic induction, and then seeded onto silk fibroin/hydroxyapatite scaffold as experimental group. Adipose-derived stem cells cultured on the cover glasses at the same condition acted as control group. The cellular morphology, proliferation and differentiation were assessed respectively by means of phase contrast microscope, MTT assay and alkaline phosphatase activity measurement.

    RESULTS AND CONCLUSION: After osteogenic induction, adipose-derived stem cells could adhere to the scaffold material and proliferate on the surface of silk fibroin/hydroxyapatite scaffold normally. No significant difference was found in cell proliferation and alkaline phosphatase activity between the experimental and control groups (P > 0.05), suggesting the cellular activity and function were not affected by the material. These findings indicate that silk fibroin/hydroxyapatite composite material has good cytocompatibility.
    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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    Bone marrow mesenchymal stem cell conditioned medium with hypoxic activation enhances its effects on radiation-induced intestinal epithelial cell injury in vitro
    Zheng Yuei, Chen Hao, Sha Wei-hong, Wang Qi-yi, Liu Wan-wei
    2015, 19 (45):  7230-7236.  doi: 10.3969/j.issn.2095-4344.2015.45.003
    Abstract ( 90 )   PDF (2120KB) ( 253 )   Save

    BACKGROUND: Conditioned medium from mesenchymal stem cells (MSC-CM) that contains abundant MSCs paracrine substances may represent a promising alternative to MSCs transplantation. However, normal MSC-CM with insufficient paracrine ability is not effective for tissue damage repair.
    OBJECTIVE: To investigate the effects of MSC-CM with (MSC-CMHyp) and without hypoxic activation (MSC-CMNor) on the proliferation and apoptosis of radiation-induced injured intestinal epithelial cells (IEC-6) and to further discuss the paracrine mechanisms.
    METHODS: IEC-6 cells were exposed to 10 Gy irradiation and cultured in MSC-CMHyp, MSC-CMNor, and DMEM-F12 medium, respectively.
    RESULTS AND CONCLUSION: Findings from trypan blue staining, flow cytometry and western blot assay showed that, compared with the DMEM-F12 medium group, treatment with MSC-CMHyp significantly enhanced IEC-6 viability proliferation after radiation-induced injury, as well as significantly decreased cell apoptosis and expression of Caspases-3/8 (P < 0.05). However, there was no significant difference between the MSC-CMNor group and DMEM-F12 medium group (P > 0.05). On the other hand, the increased levels of vascular endothelial growth factor, basic fibroblast growth factor, insulin-like growth factor-1, and interleukin-10 were detected in the MSC-CMHyp group compared to the MSC-CMNor group (P < 0.05). These results suggest that the MSC-CMHyp improves the viability and proliferative capacity of IEC-6 cells after radiation-induced injury via up-regulating secretion of cytokines and down-regulating apoptotic signaling.
    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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    Chondrogenic differentiation of bone marrow mesenchymal stem cells induced by different growth factors
    Wang Shu-ying, Lu Xiao-bing, Ouyang Lin
    2015, 19 (45):  7237-7241.  doi: 10.3969/j.issn.2095-4344.2015.45.004
    Abstract ( 101 )   PDF (1696KB) ( 246 )   Save

    BACKGROUND: Articular cartilage is a highly differentiated tissue, which is very limited in its ability to repair after injury. Stem cell therapy for cartilage repair has completely solved this problem.

    OBJECTIVE: To investigate the mechanism that different growth factors induce the differentiation of bone marrow mesenchymal stem cells into chondrocytes.
    METHODS: Passage 5 rat bone marrow mesenchymal stem cells were induced by different growth factors and their combinations, including transforming growth factor beta 1 (TGF-β1) group, TGF-β1+insulin growth factor-1 (IGF-1) group, and bone morphogenetic protein 2 (BMP-2)+IGF-1 group, TGF-β1+BMP-2 group, TGF-β1+IGF-1+ BMP-2 group, and blank control group. At 21 days of induction, cells were stained with alcian blue and alizarin red; RT-PCR was employed to detect collagen II mRNA expression.
    RESULTS AND CONCLUSION: Alcian blue staining showed metachromasia in the cytoplasm and mesenchyma, and proteoglycan was expressed green; alizarin red staining showed no orange calcium nodules. The bone marrow mesenchymal stem cells were preliminarily deduced to differentiate into chondrocytes, but could not express the cell phenotypes of bone cells. In the blank control group, the expression of collagen II mRNA was negative. Compared with the TGF-β1 group, the mRNA expression of collagen II was lower in the BMP-2+IGF-1 group, but higher in the TGF-β1+BMP-2 group and TGF-β1+IGF-1+BMP-2 group (P < 0.05). Moreover, the expression of collagen II mRNA was highest in the TGF-β1+IGF-1+BMP-2 group (P < 0.05). These findings indicate that TGF-β1 alone is able to induce the chondrogenic differentiation of bone marrow mesenchymal stem cells, and the combination of TGF-β1, IGF-1 and BMP-2 can play the biggest role to induce the chondrogenic differentiation of bone marrow mesenchymal stem cells.
    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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    Effects of allogeneic versus autologous serum on the proliferation of bone marrow stromal stem cells
    Li Fang-guo, Lu Yan-dong, Cui Meng, Zhao Jia-guo, Yang Qiang, Wang Lei, Pei Guo-xian, Sun Jie
    2015, 19 (45):  7242-7248.  doi: 10.3969/j.issn.2095-4344.2015.45.005
    Abstract ( 68 )   PDF (2448KB) ( 311 )   Save

    BACKGROUND: Fetal bovine serum as nutritional support is often used in the traditional cell culture. Consequently, a host of potential problems such as the spread of disease and immunological reactions exist. To find a suitable fetal bovine serum substitute and to establish a culture system of human bone marrow stromal stem cells in vitro which has been standardized, safe and efficient has just started.

    OBJECTIVE: To investigate the effects of different serums on proliferation of bone marrow stromal stem cells in vitro.
    METHODS: Bone marrow stromal stem cells were obtained from adult bone marrow, which were cultured in DMEM containing 10% AB serum, 10% autologous serum, or 10% fetal bovine serum. Cells at passage 3 were used in this study.

    RESULTS AND CONCLUSION: The cell confluence in the AB serum group was earlier than that in the fetal bovine serum group and autologous serum group. Human bone marrow stromal stem cells maintained the phenotypes of bone marrow stem cells in three serums detected by flow cytometry. AB serum group showed the highest fluorescence intensity and the most efficiency of cell proliferation which examined by the AlamarBlue assay. Apoptosis rate was < 5% in all the three groups, and cells grew well in these serums. Alkaline phosphatase, calcium nodules and oil red O staining showed that the cells maintained the osteogenesis and adipogenesis capacity in the three groups. AB serum was found to have a better effect on proliferation capability of cells than fetal bovine serum and autologous serum. Taken together, AB serum is expected to be a substitute of fetal bovine serum to build an in vitro culture system of adult bone marrow stromal stem cells that accord with the clinical requirements of bone tissue engineering.
    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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    Kallikrein 1-transfected bone marrow mesenchymal stem cells: selection of the multiplicity of infection
    Jia Jia, Jin Li-mei, Zhao Yi, Yan Li, Lu Juan, Wang Qiu-ping, Zhao Jing-miao, Hu Ji-hong
    2015, 19 (45):  7249-7253.  doi: 10.3969/j.issn.2095-4344.2015.45.006
    Abstract ( 82 )   PDF (2095KB) ( 288 )   Save

    BACKGROUND: Kallikrein 1 is an important component of the kallikrein-kinin system. Studies have shown that kallikrein can protect the cardiovascular system by promoting angiogenesis and inhibiting myocardial inflammation, but there is no report on its effect on inducing differentiation of stem cells.
    OBJECTIVE: To determine the transfection efficiency of kallikrein 1 adenoviral vector in rat bone mesenchymal stem cells.
    METHODS: Using adenovirus as a vector, the target gene kallikrein 1 was transfected into rat bone marrow mesenchymal stem cells. Fluorescence microscopy, MTT method and flow cytometry were used to investigate the effect of transfection and determine the optimal multiplicity of infection.
    RESULTS AND CONCLUSION: Adenovirus carrying kallikrein 1 was successfully transfected into rat bone marrow mesenchymal stem cells. Results from flow cytometry showed that the transfection efficiency was associated with the multiplicity of infection. When the multiplicity of infection was 150, the transfection efficiency was 80.8%. MTT results showed that when the multiplicity of infection was 200, the cell growth was inhibited remarkably. These findings indicate that adenovirus-mediated kallikrein 1 can be successfully transfected into rat bone marrow mesenchymal stem cells with the optimal multiplicity of infection=150.
    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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    Bone marrow mesenchymal stem cells combined with chitosan for articular cartilage injuries
    Chen Lu, Xia Xian-xue, Jiang Ke
    2015, 19 (45):  7254-7258.  doi: 10.3969/j.issn.2095-4344.2015.45.007
    Abstract ( 128 )   PDF (1949KB) ( 258 )   Save

    BACKGROUND: Trauma easily leads to the emergence of articular cartilage defects, which is a difficult problem in the orthopedics field. Tissue engineering technology provides a new method for cartilage repair.

    OBJECTIVE: To explore the feasibility of combining chitosan and bone marrow mesenchymal stem cells to repair injured articular cartilage in rabbits.
    METHODS: Cultured rabbit bone marrow mesenchymal stem cells were seeded onto chitosan scaffold, and then the composite material was implanted into the defect as experimental group. Rabbits with no treatment served as control group. Gross observation and toluidine blue staining were carried out at 6 and 12 weeks after operation.

    RESULTS AND CONCLUSION: At 6 weeks after operation, the control group had only fibrous tissue hyperplasia, and in the experimental group, cartilage-like tissues were generated at the defect site. At 12 weeks after operation, a small amount of hyaline cartilage-like tissues were observed in the control group, and the defects in the experimental group were covered with smooth and hyaline cartilage tissues. After 12 weeks, the toluidine blue staining was light in the control group with a small amount of cartilage tissues; in the experimental group, the toluidine blue staining was remarkable, and the defects were completely covered with hyaline cartilage tissues, and cartilage cells were increased in number. The findings indicate that chitosan-bone marrow mesenchymal stem cells composite material is better to induce cartilage tissue formation and promote cartilage defect repair.
    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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    Effect of astragalus polysaccharides on the neuronal differentiation of bone marrow mesenchymal stem cells
    Wang Guan, Li Jing-ya, Yang Sheng-ping, Wang Jian-min
    2015, 19 (45):  7259-7262.  doi: 10.3969/j.issn.2095-4344.2015.45.008
    Abstract ( 76 )   PDF (1510KB) ( 352 )   Save

    BACKGROUND: Active components of Astragalus have an antioxidant effect, which is considered to result in the neuron-like differentiation of bone marrow mesenchymal stem cells.
    OBJECTIVE: To investigate the effect of astragalus polysaccharides combined with basic fibroblast growth factor to induce the differentiation of bone marrow mesenchymal stem cells into nerve cells.
    METHODS: After 24 hours of pretreatment with basic fibroblast growth factor, passage 3 human bone marrow mesenchymal stem cells were cultured with astragalus polysaccharides for 1-3 days (combined group). Blank control group and basic fibroblast growth factor group were set up. Expression of neuron-specific enolase and nestin was detected using western blot or immunocytochemical staining.
    RESULTS AND CONCLUSION: The expression of neuron-specific enolase was higher in the combined group than the basic fibroblast growth factor group (P < 0.05). Expression of nestin was found in both basic fibroblast growth factor group and combined group, but the gray value was higher in the combined group than the basic fibroblast growth factor group (P < 0.05). These findings indicate that astragalus polysaccharides combined with basic fibroblast growth factor is better to induce the neuronal differentiation of bone marrow mesenchymal stem cells.
    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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    Effects of 8-bromo-7-methoxychrysin and sorafenib on apoptosis of liver cancer stem-like cells
    Aoduntuoya, Zhao Hai-zhen, Liu Rui-jun
    2015, 19 (45):  7263-7267.  doi: 10.3969/j.issn.2095-4344.2015.45.009
    Abstract ( 76 )   PDF (1719KB) ( 412 )   Save

    BACKGROUND: We tried to combine 8-bromo-7-methoxychrysin and sorafenib in order to offset the tolerance of hepatocellular cancer stem cells to sorafenib, thereby comprehensively improving the therapeutic efficacy on hepatocellular carcinoma.
    OBJECTIVE: To observe the effects of 8-bromo-7-methoxychrysin, sorafenib and their combination on apoptosis of liver cancer stem-like cells SMMC-7721, and to analyze their mechanisms.
    METHODS: SMMC-7721 cells were treated with 8-bromo-7-methoxychrysin, sorafenib alone and their combination for 24 hours. Then, flow cytometry was used to detect cell apoptosis and western blot assay was used to determine nuclear factor-κB protein expression.
    RESULTS AND CONCLUSION: Compared with 8-bromo-7-methoxychrysin group and sorafenib group, the apoptotic rates of SMMC-7721 cells were significantly enhanced after treatment with the combination of 8-bromo-7-methoxychrysin group and sorafenib (2.5, 5, 10, 50 μmol/L), and meanwhile, the protein expression of nuclear factor-κB was down-regulated significantly. These findings indicate that the combined therapy of 8-bromo-7-methoxychrysin group and sorafenib can enhance the apoptosis of SMMC-7721 cells, which may beassociated with down-regulation of nuclear factor-κB protein.
    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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    Effect of cobalt chloride-induced hypoxia on proliferation of human umbilical cord-derived mesenchymal stem cells and related gene and protein expressions
    Han Xiao, Bai Hai, Yin Jiao-jiao, Yang Ke, Han Yan-xia, Ou Jian-feng, Wang Cun-bang
    2015, 19 (45):  7268-7273.  doi: 10.3969/j.issn.2095-4344.2015.45.010
    Abstract ( 95 )   PDF (2304KB) ( 527 )   Save

    BACKGROUND: Cobalt chloride (CoCl2) may promote the proliferation of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) in a time- and concentration-dependent manner, and meanwhile, CoCl2 can regulate the expression of genes and proteins in hUC-MSCs.
    OBJECTIVE: To explore the effects of CoCl2 induced-hypoxia on the proliferation of hUC-MSCs and gene and protein expressions in hUC-MSCs, thereby establishing an effective method for MSCs culture and amplification in vitro.
    METHODS: hUC-MSCs were extracted using tissue explant method. Under hypoxia conditions induced by CoCl2 (0, 100, 150, 200, 250 μmol/L) for different periods (0, 1, 2, 3, 4 days), flow cytometry was used to identify cell surface-associated antigens; cell counting kit-8 was used to detect cell proliferation; RT-PCR was used to 
    determine levels of hypoxia inducible factor-1α, inducible nitric oxide synthase, stromal cell-derived factor-1, interleukin-6, transforming growth factor-β mRNA; western blot assay was used to detect protein expression of hypoxia inducible factor-1α.
    RESULTS AND CONCLUSION: The cells were positive for CD29, CD73, CD90, CD105, while negative for CD31, CD14, CD34, CD45, CD11b, HLA-DR. Moreover, the antigen expression was not affected by CoCl2 induced-hypoxia. CoCl2 induced-chemical hypoxia could promote the proliferation of hUC-MSCs in a time- and concentration-dependent manner. RT-PCR results showed that under hypoxia, hypoxia inducible factor 1α, inducible nitric oxide synthase and stromal cell-derived factor-1 mRNA expressions were significantly up-regulated, but interleukin-6 and transforming growth factor-β mRNA expressions were down-regulated significantly (P < 0.05). Additionally, the protein expression of hypoxia inducible factor 1α was increased under hypoxia conditions. These findings indicate that CoCl2 induced-hypoxia environment may promote the proliferation of hUC-MSCs and the optimal concentration of CoCl2 is 200 μmol/L. However, a higher concentration of CoCl2 (≥ 250 μmol/L) inhibits the proliferation of hUC-MSCs, and the mechanism may be related to the increase of hypoxia inducible factor-1α at protein and mRNA levels.
    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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    Effect of human umbilical cord mesenchymal stem cells on the proliferative characteristics of cervical cancer Hela cells
    2015, 19 (45):  7274-7278.  doi: 10.3969/j.issn.2095-4344.2015.45.011
    Abstract ( 124 )   PDF (1969KB) ( 245 )   Save

    BACKGROUND: Mesenchymal stem cells have the specific chemotaxis to the inflammation and tumor tissue, but the effect of mesenchymal stem cells on the growth of cervical cancer cells becomes an urgent problem.
    OBJECTIVE: To investigate the effect of human umbilical cord mesenchymal stem cells on the proliferation of cervical cancer Hela cells.
    METHODS: Hela cells were co-cultured with human umbilical cord mesenchymal stem cells at different number or its conditioned medium at different concentrations for 3 days. Then, cell counting kit-8 was used to detect the proliferation of Hela cells.
    RESULTS AND CONCLUSION: When Hela cells were co-cultured with human umbilical cord mesenchymal stem cells at ratios of 1:3, 1:2, 1:1, 2:1, 4:1, 8:1, the relative proliferation inhibition rates were 67.12%, 47.18%, 31.15%, 27.61%, 15.55% and 15.95%, respectively. When the Hela cells were co-cultured with human umbilical cord mesenchymal stem cell conditioned medium at 5, 10, 20, 40, 60, 100 mg/L, the relative proliferation inhibition rates were 0.61%, 40.1%, 63.47%, 80.61%, 93.56%, 90.65%, respectively. These findings indicate that the proliferation of Hela cells can be inhibited by co-culture with human umbilical cord mesenchymal stem cells at a certain concentration-dependent manner.
    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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    Fibroblast growth factor-modified bone marrow mesenchymal stem cells promote functional recovery from traumatic brain injury
    Li Xue-dong, Chen Jia-kang, Qin Jun, Mai Yong-jun, Xiao Zhen-yong
    2015, 19 (45):  7279-7285.  doi: 10.3969/j.issn.2095-4344.2015.45.012
    Abstract ( 72 )   PDF (1046KB) ( 275 )   Save
    BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) can promote nerve regeneration, but there are no better results because of the limitations of treatment methods. BMSC transplantation alone is not enough to achieve desired therapeutic effects.
    OBJECTIVE: To investigate the effect of fibroblast growth factor (FGF)-modified BMSC transplantation on functional recovery and expression of glial fibrillary acidic protein after traumatic brain injury.
    METHODS: Animal models of traumatic brain injury were established in Sprague-Dawley rats using hydraulic shock method, and then randomized into control group (traumatic brain injury group), BMSC group and FGF-BMSC group (FGF-modified BMSC group). After isolation and culture, BMSCs were modified by adenovirus vector-mediated FGF gene. Western blot assay was used to detect transfection efficiency and glial fibrillary acidic  
    protein expression; immunohistochemical detection was used to detect distribution and number of BrdU positive cells in the brain; Longa score was used to evaluate the neurologic function of rats at 1, 3 days, 1, 2 weeks after transplantation; TUNEL assay was used to detect cell apoptosis in the brain.
    RESULTS AND CONCLUSION: Western blot results showed that FGF gene was successfully transferred to the adenovirus vector, and capable of expressing in BMSCs; moreover, the glial fibrillary acidic protein expression of FGF-BMSC group was significantly higher than that in the other two groups (P < 0.05). The number of BrdU positive cells in the brain was significantly higher in the FGF-BMSC group than the other two groups (P < 0.05). Two weeks after transplantation, the Longa scores in the FGF-BMSC group were significantly lower than those in the other two groups (P < 0.05). TUNEL results showed that the number of apoptotic cells in the FGF-BMSC group was significantly lower than that in the other two groups (P < 0.05). These findings indicate that FGF-modified BMSCs transplantation is able to improve neurological damage after traumatic brain injury and promote neurological recovery, which is better than BMSC transplantation alone. 
    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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    Protein expression and biological characteristics of colon cancer stem cells: culture and screening
    Zhang Dong-xing, Yan Deng-guo
    2015, 19 (45):  7286-7291.  doi: 10.3969/j.issn.2095-4344.2015.45.013
    Abstract ( 129 )   PDF (1006KB) ( 315 )   Save

    BACKGROUND: EpCAMhighCD44+ colon cancer stem cells have been isolated in recent years, and most of them are related to the study of isolating tumor stem cells in colorectal cancer cell lines. There are less reports on the function of colon cancer stem cells.
    OBJECTIVE: To isolate, screen and culture colon cancer stem cells from colorectal carcinoma tissues and to explore the protein expression and biological characteristics of colon cancer stem cells.
    METHODS: The primary cells were isolated from the colon cancer tissues. EpCAMhighCD44+ cells were detected and sorted by flow cytometry. Then, these cells were cultured in serum-free medium. After the cells were 
    replanted subcutaneously into the mice, we observed the daily performance and tumor growth in the mice. When the tumor diameter was above 1 cm, tumor tissues were taken for immunohistochemical testing. The growth of EpCAMhighCD44+ cells cultured in the serum-free medium was observed; expressions of neutral epithelial mucin and CK-20 in EpCAMhighCD44+ cells were detected.
    RESULTS AND CONCLUSION: EpCAMhighCD44+ colon cancer stem cells were detected in the colon cancer tissues, and the proportion of EpCAMhighCD44+ cells in primary colon cancer cells was 1.7%-38%. After 24 hours of serum-free culture, there were a few of small suspended cell spheres which were incompact. After 21 days, dense cell spheres with uniform size were found. The formation of transplanted tumor was found in the nude mice. Hematoxylin-eosin staining showed the transplanted tumor had the typical characteristics of colon cancer cells. The expression of neutral epithelial mucin and CK-20 was observed in all the transplanted tumors. Additionally, the proliferation of EpCAMhighCD44+ cells could be promoted by 5-fluorouracil (10, 1, and 0.1 PPC) with time. These findings indicate that EpCAMhighCD44+ colon cancer stem cells in the colon cancer have a certain ability of proliferation, regeneration, differentiation, tumor formation and chemotherapy resistance.
    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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    Ginsenosides-induced bone marrow mesenchymal stem cells promote nerve regeneration in traumatic brain injury
    Qin Jun, Chen Jia-kang, Li Xue-dong, Mai Yong-jun, Xiao Zhen-yong
    2015, 19 (45):  7292-7297.  doi: 10.3969/j.issn.2095-4344.2015.45.014
    Abstract ( 80 )   PDF (861KB) ( 461 )   Save

    BACKGROUND: Previous studies have shown that bone marrow mesenchymal stem cells in the treatment of neurological diseases have achieved some success, which can promote neurological alterations; however, there is no breakthrough on gene and drug regulation.
    OBJECTIVE: To investigate the influence of ginsenosides-induced differentiation of bone marrow mesenchymal stem cells on nerve regeneration after traumatic brain injury.
    METHODS: A traumatic brain injury model was built in rats using hydraulic shock method, and then rat models were randomly divided into model group (traumatic brain injury group), bone marrow mesenchymal stem cell group, ginsenosides group (ginsenosides induced differentiation of bone marrow mesenchymal stem cells). At 2 weeks after transplantation, western blot assay was used to detect protein expression levels of nerve growth factor and brain-derived neurotrophic factor, immunohistochemistry assay used to detect the number of BrdU-positive cells. At 1, 3 days and 1, 2 weeks after transplantation, modified neurological severity scores were recorded.
    RESULTS AND CONCLUSION: The expression levels of nerve growth factor and brain-derived neurotrophic factor protein were significantly higher in the ginsenosides group than the bone marrow mesenchymal stem cell 
    group and model group (P < 0.05). The number of BrdU positive nerve cells was also higher in the ginsenosides group than the bone marrow mesenchymal stem cell group and model group (P < 0.05). At 3 days and 1, 2 weeks after transplantation, the modified neurological severity scores in the ginsenosides group were lower than those in the bone marrow mesenchymal stem cell group and model group (P < 0.05). These findings indicate that ginsenoside-induced bone marrow mesenchymal stem cell transplantation can promote nerve regeneration in rats with traumatic brain injury, which has better outcomes than bone marrow mesenchymal stem cell transplantation alone.
    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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    Therapeutic effect of umbilical cord-derived mesenchymal stem cell transplantation in systemic lupus erythematosus patients with different patterns of syndromes
    Tang Yu, Liu Rui-xia, Qiu Ying-ying, Rui Jin-bing, Li Jing
    2015, 19 (45):  7298-7303.  doi: 10.3969/j.issn.2095-4344.2015.45.015
    Abstract ( 88 )   PDF (910KB) ( 281 )   Save

    BACKGROUND: Systemic lupus erythematosus (SLE) is classified into four types, and the major treatment is to tonify kidney and nourish yin, clear blood stasis and toxin by the traditional Chinese medicine (TCM). Even though, there are still many patients with poor efficacy. Mesenchymal stem cells have the capacity of multiple differentiation, hematopoietic support and immune regulation, thus having been used for the treatment of refractory, recurrent SLE and achieving good effects.
    OBJECTIVE: To investigate the therapeutic effect of umbilical cord-derived mesenchymal stem cell transplantation on SLE patients with different patterns of syndromes.
    METHODS: Twenty-one SLE patients were clustered to four syndrome types of TCM, including heat-toxin, yin deficiency of liver and kidney, yang deficiency of spleen and kidney, and qi stagnation and blood stasis. The changes in clinical and laboratory indicators were analyzed statistically before and after cell transplantation.
    RESULTS AND CONCLUSION: The level of 24-hour proteinuria and SLE disease activity index scores in SLE patients were significantly decreased at 1, 3, 6 months after cell transplantation (P < 0.01). Umbilical cord-derived mesenchymal stem cell transplantation could significantly reduce the 24-hour proteinuria in SLE patients with yin deficiency of liver and kidney at 1, 3 and 6 months (P < 0.01), while slightly reduce the 24-hour proteinuria in SLE patients with heat-toxin and qi stagnation and blood stasis at 1, 3 months (P < 0.05) as well as in SLE patients 
    with yang deficiency of spleen and kidney at 1 month (P < 0.05). Additionally, umbilical cord-derived mesenchymal stem cell transplantation could increase the serum albumin levels in all the SLE patients (P < 0.01), although the changes in patients with heat-toxin were moderate (P < 0.05). All the SLE patients of four types had an increasing trend of their platelet counting after cell transplantation, but there was no statistical difference before and after cell transplantation. Taken together, umbilical cord-derived mesenchymal stem cell transplantation is effective for treatment of SLE, but has different therapeutic efficacy on SLE patients with different syndrome types of TCM. 
    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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    Homing of human umbilical cord-derived mesenchymal stem cells to the injured kidney and their protective effects
    Zhang Bing, Li Wen-qing
    2015, 19 (45):  7304-7308.  doi: 10.3969/j.issn.2095-4344.2015.45.016
    Abstract ( 110 )   PDF (766KB) ( 489 )   Save

    BACKGROUND: An increasing number of studies have shown that mesenchymal stem cells have the potential to treat acute kidney injury. Umbilical cord-derived mesenchymal stem cells have general characteristics of stem cells and many advantages, such as easy to isolate and culture, in vitro fast amplification, low immunogenicity and no ethical problems, which have garnered increasing attentions.
    OBJECTIVE: To study the repairing effects of human umbilical cord-derived mesenchymal stem cells on acute kidney injury in rats.
    METHODS: Thirty rats were randomized into three groups: a normal control group, a model group and a cell transplantation group. Rats in the model and cell transplantation were subjected to clamping the renal pedicles for 45 minutes, and then injected 1 mL of DAPI-labeled umbilical cord-derived mesenchymal stem cells or 1 mL of saline via the tail vein. In the normal control group, the kidney was only exposed with no treatment. At 7 days after treatment, the rats were killed to take left kidney tissues for pathological observation under light microscope and right kidney for observation of DAPI-positive cell counting. Automatic biochemical analyzer was used to detect serum creatinine and urea ammonia levels.
    RESULTS AND CONCLUSION: Compared with the model group, the levels of serum creatinine and urea ammonia were significantly lower in the cell transplantation group (P < 0.05), suggesting that human umbilical cord-derived mesenchymal stem cells can improve the kidney function to a certain extent. Pathological findings 
    showed that the pathological damage was improved more remarkably in the cell transplantation group than the model group, and the tubular necrosis index decreased significantly in the cell transplantation group. At 7 days after cell transplantation, blue fluorescent cells were scattered on renal tissue frozen sections. These results indicate that human umbilical cord-derived mesenchymal stem cells can migrate to the injured tubular epithelial tissues, and promote the repair of the injured kidney. 
    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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    Combined therapy of acupuncture and neural stem cell transplantation in immature rats with cerebral palsy
    Jie Xiao-su, Hou Yu-jin
    2015, 19 (45):  7309-7313.  doi: 10.3969/j.issn.2095-4344.2015.45.017
    Abstract ( 72 )   PDF (1951KB) ( 328 )   Save

    BACKGROUND: Both acupuncture and neural stem cell therapy have good achievements in the treatment of cerebral palsy, but there are few reports about their combined therapy.
    OBJECTIVE: To observe the therapeutic effect of combined therapy of acupuncture and neural stem cell transplantation on cerebral palsy rats and to explore the relevant mechanism of action.
    METHODS: Cerebral palsy models were built in newborn rats, and then, the model rats were subjected to acupuncture, neural stem cell transplantation and their combination, respectively. Meanwhile, normal control group and model group were set up.
    RESULTS AND CONCLUSION: Compared with the other groups, in the combined therapy group, the behavior scores of cerebral palsy rats improved significantly (P < 0.05), the number of nerve cells and Bcl-2 protein expression were increased significantly (P < 0.05), and the number of apoptotic nerve cells and Bax protein expression were decreased significantly (P < 0.05). Additionally, there were no significant differences in these indicators among the model, acupuncture and neural stem cell transplantation groups. These findings indicate that the combined therapy of acupuncture and neural stem cell therapy can obviously improve learning and memory ability of cerebral palsy rats and reduce neuronal apoptosis, which is probably associated with down-regulation of Bax protein and up-regulation of Bcl-2 protein. 
    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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    Neural stem cell transplantation for partial sciatic nerve transaction-induced neuropathic pain: the optimal cell number for transplantation
    Deng Mo, Zhao Feng
    2015, 19 (45):  7314-7319.  doi: 10.3969/j.issn.2095-4344.2015.45.018
    Abstract ( 86 )   PDF (1889KB) ( 352 )   Save

    BACKGROUND: Studies have shown that neural stem cell transplantation has a certain effect on neuropathic pain, but the efficacy of transplanted cell number on neuropathic pain is not exactly understood.
    OBJECTIVE: To observe the effect of different amount of neural stem cells administered intrathecally on the neuropathic pain and expression of glial-derived neurotrophic factor in the spinal cord dorsal horn and dorsal root ganglion in rats after partial sciatic nerve transaction.
    METHODS: A Sprague-Dawley rat at 14-16 days of pregnancy was used to prepare neural stem cell suspensions that were injected intrathecally into rat models of partial sciatic nerve transaction at doses of 1×103, 1×104, 1×105, 1×106, 1×107 cells per 30 μL, respectively. Additionally, model group and sham-operated group were set up. Threshold values of mechanical and thermal pain were recorded 1 day before operation, 1, 3, 7, 14, 
    21 days after operation. Expressions of glial-derived neurotrophic factor protein and mRNA in the ipsilateral spinal cord dorsal horn and dorsal root ganglion were detected by immunohistochemistry and RT-PCR, respectively, at 7 and 21 days after partial sciatic nerve transaction.
    RESULTS AND CONCLUSION: Pain threshold values were decreased in all the groups except the sham-operated group at 1 day after operation, and reached the peak at 7 and 14 days after operation (P < 0.05). Compared with the model group, the pain threshold values and the expression of glial-derived neurotrophic factor protein and mRNA in the ipsilateral spinal cord dorsal horn and dorsal root ganglion were increased gradually in a dose-dependent manner in the 1×104, 1×105, 1×106, 1×107 groups at 7 days after operation (P < 0.05). At 21 days post-operation, the pain threshold values showed no differences from the preoperative findings in the 1×105, 1×106, 1×107 groups, but the expression of glial-derived neurotrophic factor was significantly higher in the 1×105 group than the other groups (P < 0.05). Taken together, intrathecal transplantation of neural stem cells at a dose of 1×105 is the most effective in alleviating partial sciatic nerve transaction-induced neuropathic pain in rats. 
    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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    Combining bone marrow mesenchymal stem cells with new nerve conduits for repair of peripheral nerve defects
    Zhu De-zhi, Wang Bo-min
    2015, 19 (45):  7320-7324.  doi: 10.3969/j.issn.2095-4344.2015.45.019
    Abstract ( 85 )   PDF (2157KB) ( 189 )   Save

    BACKGROUND: With the continuous development and progress of medical model, the treatment and rehabilitation recovery from peripheral nerve defects needs higher requirements; therefore, stem cell culture-based nerve tissue engineering technology provides a new strategy for nerve defect repair.
    OBJECTIVE: To explore the feasibility of combining bone marrow mesenchymal stem cells and new nerve conduits to repair peripheral nerve defects.
    METHODS: Fifty New Zealand white rabbits, clean grade, were randomly divided into experimental group and control group, with 25 rabbits in each group. Animal models of peripheral nerve defects were made about 15 mm distant to the middle segment of the radius of the rabbit foreleg. At 1 week after modeling, bone marrow mesenchymal stem cells/new nerve conduit composite material was implanted into the defect in the experimental group; and in the control group, only bone marrow mesenchymal stem cells were transplantation. At 4 weeks after transplantation, a 5-mm nerve fiber was taken from the defect site, stained with hematoxylin-eosin and observed under scanning electron microscope. Density and diameter of regenerated nerve fibers were compared between the two groups.
    RESULTS AND CONCLUSION: Compared with the control group, the density of nerve fibers was significantly higher in the experimental group, but the diameter of nerve fibers was significantly lower (P < 0.05). There were more cells with good growth, large size and many processes on the surface of regenerated nerve tissues in the experimental group than the control group. Moreover, in the experimental group, interconnected cells were woven into a mesh and the axons were longer and thicker, both of which were the performance of typical neuron-like cells. Taken together, the bone marrow mesenchymal stem cells/new nerve conduit composite material can be used for peripheral nerve repair and present exact achievements. 
    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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    Bone marrow mesenchymal stem cell transplantion for repair of airway injury in chronic obstructive pulmonary disease
    Zhang Qi, Li Dan
    2015, 19 (45):  7325-7330.  doi: 10.3969/j.issn.2095-4344.2015.45.020
    Abstract ( 80 )   PDF (2460KB) ( 313 )   Save
    BACKGROUND: Mesenchymal stem cells can differentiate into lung parenchymal cells involved in lung injury repair, providing a new approach for the application of mesenchymal stem cells in patients with chronic obstructive pulmonary disease.
    OBJECTIVE: To observe the effect of bone marrow mesenchymal stem cell transplantation on the repair of airway injury in rats with chronic obstructive pulmonary disease.
    METHODS: Twenty-four female rats were randomized into four groups: bone marrow mesenchymal stem cell transplantation group (cell transplantation group, n=12); bone marrow mesenchymal stem cells group (cell control 
    group, n=4); model group (n=4); healthy control group (n=4). Rat models of chronic obstructive pulmonary disease were established in the cell transplantation group and model group using fumigation+lipopolysaccharide method; and at 1 day after modeling, model rats were given 1 mL CM-Dil-labeled bone marrow mesenchymal stem cells and 1 mL PBS via the tail vein in these two groups, respectively. In addition to tracheal injection of normal saline (300 μL) at 1 and 14 days, rats in the cell control and healthy control groups were given 1 mL CM-Dil-labeled bone marrow mesenchymal stem cells and 1 mL PBS via the tail vein, respectively. At 1, 7, 15 and 30 days after cell transplantation, lung tissue and serum markers of all rats were detected.
    RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining showed that emphysema and airway injury was milder in the cell transplantation group than the model group, but severer than the cell control and healthy control groups. (2) The total number of leukocytes and neutrophils in the peripheral blood was higher in the cell transplantation group than the cell control and healthy control groups (P < 0.05); with time, the total number of leukocytes and neutrophils was decreased gradually. (3) Compared with the cell control and healthy control groups, the interleukin-10 level in the peripheral blood was lower and the levels of tumor necrosis factor-α and granulocyte colony-stimulating factor were higher at 1 day after cell transplantation (P < 0.05). With time, in the cell transplantation group, the interleukin-10 level was increased gradually, the level of tumor necrosis factor-α was decreased gradually, and the level of granulocyte colony-stimulating factor was increased first and then decreased, which was highest at 7 days after cell transplantation. (4) Partial CM-Dil-positive cells were positive for CC16. Taken together, bone marrow mesenchymal stem cell transplantation via the tail vein can improve lung injury of rats with chronic obstructive pulmonary disease, and it is involved in the repair of airway injury through differentiation into epithelial cells and immune regulation.
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    Neuroprotective effect of neural stem cells modified by glial-derived neurotrophic factor on cerebral apoplexy
    Wu Zhong-hua
    2015, 19 (45):  7331-7336.  doi: 10.3969/j.issn.2095-4344.2015.45.021
    Abstract ( 65 )   PDF (801KB) ( 269 )   Save

    BACKGROUND: Glial-derived neurotrophic factor has a specific effect on brain neurons, and is an important neurotrophic factor in the treatment of cerebral apoplexy.
    OBJECTIVE: To investigate the neuroprotective effect of glial-derived neurotrophic factor gene modified neural stem cells on rat cerebral apoplexy.
    METHODS: The recombinant human plasmid pAdEasy-l-pAdTrackCMV was constructed, and the neural stem cells were isolated and cultured from the cortex of neonatal rats. The neural stem cells were transfected with the recombinant adenovirus of glial-derived neurotrophic factor, and the cell suspension was injected into the right brain ventricle of rats with transient cerebral ischemia (2 hours). Meanwhile, neural stem cell transplantation group and control group were set up.
    RESULTS AND CONCLUSION: Compared with the neural stem cell transplantation group, the modified neurological severity score of combined transplantation group was reduced significantly 2 and 3 weeks after reperfusion, and the area of cerebral ischemia injury was also significantly decreased at 7 days after reperfusion (P < 0.05). The number of neural stem cells in the neural stem cell transplantation group was significantly less than that in the combined transplantation group (P < 0.05). The expression of Syn, PSD-95 proteins in the two transplantation groups, especially in the combined transplantation group, was higher than that of the control group (P < 0.05). However, there was no significant difference between the two transplantation groups (P > 0.05). The results show that the neural stem cells modified by glial-derived neurotrophic factor can play a better role in the neuroprotection against cerebral apoplexy in rats, and the effect is better than that of simple neural stem cells.
    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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    Differentiation of neural stem cells induced by erythropoietin in vitro
    Zhao Yan, Wang Xi-liang, Xiao Yu-long, Huo Hong-jun, Jiang Jian-ming, Yan Hui-bo
    2015, 19 (45):  7337-7341.  doi: 10.3969/j.issn.2095-4344.2015.45.022
    Abstract ( 142 )   PDF (1416KB) ( 216 )   Save

    BACKGROUND: In recent years, neural stem cells are considered to be ideal for the treatment of spinal cord injury, but the proportion of its natural differentiation into neurons in the host body is relatively low, which severely restricts the therapeutic effect on spinal cord injury.
    OBJECTIVE: To investigate the effect of erythropoietin on the differentiation of neural stem cells in vitro.
    METHODS: Under sterile condition, neural stem cells from the hippocampus of neonatal Wistar rats were isolated, cultured and identified by immunofluorescence in vitro. The third generation of neural stem cells were randomly divided into 0.5, 5, 50 U/mL erythropoietin groups and control group (with no erythropoietin).
    RESULTS AND CONCLUSION: Compared with the control group, the differentiation rate of neural stem cells was significantly improved in the 0.5, 5, 50 U/mL erythropoietin groups (P < 0.05); moreover, the differentiation rate of neural stem cells in the 5, 50 U/mL erythropoietin groups was higher than that in the 0.5 U/mL erythropoietin group (P < 0.05). In addition, there was no significant difference between the 5, 50 U/mL erythropoietin groups (P > 0.05). These findings indicate that erythropoietin can effectively induce the  differentiation of neural stem cells into neurons in vitro, and moreover, it can significantly improve the differentiation rate of neural stem cells into neurons.
    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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    Effects of amniotic fluid stem cell transplantation on immune tolerance and oxidative stress in kidney transplantation
    Deng Chun-yang, Feng Jian-xun, Zhang Hai-ying, Chen Ting-fang, Li Jing
    2015, 19 (45):  7342-7349.  doi: 10.3969/j.issn.2095-4344.2015.45.023
    Abstract ( 121 )   PDF (1152KB) ( 359 )   Save

    BACKGROUND: Stem cells can induce immune tolerance, prolong graft survival time and reduce rejection in organ transplantation, which have become a hot research.
    OBJECTIVE: To induce immune tolerance to allogenic kidney transplantation with amniotic fluid stem cells in recipient rats and to explore the mechanism underlying immune tolerance.
    METHODS: Amniotic fluid stem cells were isolated from Wistar rats. Two inbred male rat strains, Wistar rats and Sprague-Dawley rats, were selected as donors and recipients of kidney transplantation. The rat models of renal orthotopic transplantation were divided into the following four groups: a sham-operated group (n=10, Sprague-Dawley rats); an isograft group (n=10, Sprague-Dawley to Sprague-Dawley rats); a control group (n=10, Wistar to Sprague-Dawley rats, treated with 1 mL saline); and an experimental group (n=10, Wistar to Sprague-Dawley rats, treated with 1 mL of 3×106/L amniotic fluid stem cells). Serum levels of creatinine, urea 
    nitrogen, interleukin-2, interferon-γ, parameters of oxidative stress were detected at 5 days after operation. Flow cytometry was employed to determine the percentage of CD4+ and CD8+ lymphocytes in the peripheral blood. Kidney transplants were observed pathologically.
    RESULTS AND CONCLUSION: Compared with the control group, the levels of creatinine, urea nitrogen, interleukin-2, interferon-γ, parameters of oxidative stress and proteinuria were lower in the experimental group (P < 0.05). Percentages of CD4+, CD8+ and CD4+/CD8+ ratio were also significantly lower in the experimental group than the control group. However, the rate of cretinemia clearance in the experimental group was significantly higher than that in the control group (P < 0.05). Furthermore, the degree of kidney injury in the experimental group was significantly lower than that in the control group. Our findings demonstrate that the amniotic fluid stem cell transplantation can induce immune tolerance, extenuate oxidative stress, attenuate pathological damage to the kidney transplant and preserve kidney function from acute rejection in rats undergoing kidney transplantation.
    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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    Design of a centrifuge device for high acceleration loading on cells
    Ge Hong-yu, Zhang Chun-qiu, Gao Li-lan, Zhang Xi-zheng
    2015, 19 (45):  7350-7355.  doi: 10.3969/j.issn.2095-4344.2015.45.024
    Abstract ( 119 )   PDF (2282KB) ( 676 )   Save

    BACKGROUND: With the development of science and technology and modern aerospace, the effects of mechanobiology in extreme mechanical environment—high acceleration are becoming an issue of concern. Studies have shown that high acceleration has certain effects on the cells.
    OBJECTIVE: Based on a centrifuge, to design a cell loading centrifuge used for exploration of cell mechanobiology under high acceleration.
    METHODS: For the cell loading centrifuge, a culture plate or/and culture bottle full of culture fluid was/were loaded with constant acceleration or variable velocity to explore the experimental feasibility. Besides, a finite element model was built by ANSYS software according to structure and properties of the rotor. The rotor system was calculated under equilibrium and dangerous working conditions, respectively, to analyze the stress and deformation distribution. Moreover, the strength of the main shaft was checked under the dangerous working conditions. Then the analysis results of ANSYS were compared with the results of strength check.
    RESULTS AND CONCLUSION: Experimental findings showed that the culture plate or/and the culture bottle could be used for (0-40)×g highly constant acceleration or variable acceleration loading. Through the simulation and comparison analysis, we confirmed the reliability of the cell loading centrifuge. This cell loading centrifuge can be used to implement the study of cell mechanobiology under high acceleration in the general biology laboratory. It also provides a basis for wide application of cell loading centrifuge in the future.
    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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    TLR2 expression in peripheral blood mononuclear cells of Henoeh-Schonlein purpura children and its association with immune response
    Zhang Zi-li, Wang Gao-feng, Mei Dao-qi, Lin Peng-fei, Tian Ling
    2015, 19 (45):  7356-7361.  doi: 10.3969/j.issn.2095-4344.2015.45.025
    Abstract ( 64 )   PDF (825KB) ( 318 )   Save

    BACKGROUND: Toll-like receptor (TLR) and its signaling pathway play an important role in autoimmune diseases, hypersensitivity, inflammation, apoptosis and transplant rejection; however, its effects on immune pathogenesis of Henoeh-Schonlein purpura in children have not been fully elucidated.

    OBJECTIVE: To investigate the TLR2 expression in peripheral blood mononuclear cells in children with Henoeh-Schonlein purpura and its correlation with immune response.
    METHODS: Sixty-four children with Henoeh-Schonlein purpura were divided into two groups: non-renal damage group (n=36) and renal damage group (n=28). Meanwhile, another 30 healthy children subjected to health examination acted as control group. Flow cytometry and florescent quantitative PCR were employed to detect TLR2 protein and mRNA expression in peripheral blood mononuclear cells, respectively. ELISA was used to detect plasma interferon-γ and interleukin-4 levels and transforming growth factor β and interleukin-10 levels secreted from Treg cells.
    RESULTS AND CONCLUSION: Levels of interferon-γ and interferon-γ/interleukin-4 in the children with Henoeh-Schonlein purpura were significantly lower than those in the control group (P < 0.05), while the level of interleukin-4 was higher than the control group (P < 0.05). The expression of TLR2 protein and mRNA was significantly higher in the Henoeh-Schonlein purpura children than the healthy children (P < 0.05) and significantly higher in the renal damage group than the non-renal damage group (P < 0.05). Compared with the control group, the levels of interleukin-10 and transforming growth factor β were significantly higher in the children with Henoeh- Schonlein purpura (P < 0.05). These findings indicate that Henoeh-Schonlein purpura children have increased levels of TLR2 protein and mRNA in the peripheral blood mononuclear cells, and exhibit immune imbalance. TLR2 is involved in the pathogenesis of Henoeh-Schonlein purpura, and transforming growth factor β can be used to evaluate Treg immune response and provide reference for diagnosis, treatment of prognosis of Henoeh-Schonlein purpura children.
    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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    Mesenchymal stem cells in the inflammatory immunomodulation 
    Zheng Sheng, Yang Juan, Tang Ying-mei
    2015, 19 (45):  7362-7368.  doi: 10.3969/j.issn.2095-4344.2015.45.026
    Abstract ( 125 )   PDF (721KB) ( 543 )   Save
    BACKGROUND: Studies have shown that the main functions of mesenchymal stem cells include direct participation in wound healing, growth factor secretion, promoting angiogenesis, immune regulation and inflammation, anti-oxidative stress, which can be used to treat a variety of acute and chronic diseases.
    OBJECTIVE: To review advances in mesenchymal stem cells in the inflammatory immunomodulation.
    METHODS: A computer-based search of Wanfang, CNKI and PubMed databases was performed for articles concerning advances in mesenchymal stem cells in the inflammatory immunomodulation published from January 2005 to August 2015. The search terms were “stem cells, mesenchymal stem cells, immune regulation, inflammation, immune cells, inflammatory factors, treatment” in Chinese and English, respectively. Finally, 40 articles were included in result analysis.
    RESULTS AND CONCLUSION: Because of their immunomodulation and muti-directional differentiation, mesenchymal stem cells garner increasing attentions. In addition, mesenchymal stem cells can be harvested from different tissues and have good in vitro amplification capability, which have a broad prospect in the clinical use, including tissue repair and anti-inflammation. As the most promising cells used clinically, mesenchymal stem cells show their superiority in the treatment of many diseases, especially in inflammations induced by immune modulation imbalance. We believe that mesenchymal stem cells will play an important role in the future cell biotherapy.
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    Periodontal ligament stem cell differentiation and proliferation in periodontal tissue regeneration
    Zhu Ling-lan, Xu Sheng-min
    2015, 19 (45):  7369-7373.  doi: 10.3969/j.issn.2095-4344.2015.45.027
    Abstract ( 121 )   PDF (640KB) ( 282 )   Save

    BACKGROUND: Periodontal ligament stem cells are one of the ideal seed cells in periodontal tissue regeneration. Sources, biological characteristics and influential factors of periodontal ligament stem cells have been an issue of concern.
    OBJECTIVE: To review the biological characteristics and functional factors of periodontal ligament stem cells as well as relevant research status and progress in regenerative medicine, and to discuss the relevant application prospect and existing problems, thereby providing theoretical and experimental basis.
    METHODS: PubMed and Wanfang databases were searched by the first author for articles related to differentiation and proliferation of periodontal ligament stem cells published from 2002 to 2015. The key words were “periodontal ligament stem cell, periodontal tissue, proliferation, differentiation” in English and Chinese, respectively. Finally, 47 articles were included in result analysis.
    RESULTS AND CONCLUSION: Periodontal ligament stem cells have the ability of differentiating into fibroblasts, adipocytes, chondrocytes and cementum. Cell growth factors play an important role in the proliferation and differentiation of periodontal ligament stem cells. Fibroblast growth factor, vascular endothelial growth factor and insulin-like growth factor can promote the proliferation of periodontal ligament stem cells. Application of stem cell biofilm as a carrier material can effectively guide periodontal tissue regeneration. Isolation, culture and influential factors of periodontal ligament stem cells should be further improved. Combined application of stem cells, biofilms and growth factors is expected to achieve the desired periodontal tissue regeneration, which is the focus of future research.
    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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    Extracorporeal shock wave and myocardial angiogenesis: effects on endogenous stem cells, cytokines and local microenvironment 
    Ma Yi-ming, Li Li, Cai Hong-yan, Guo Tao
    2015, 19 (45):  7374-7380.  doi: 10.3969/j.issn.2095-4344.2015.45.028
    Abstract ( 72 )   PDF (926KB) ( 261 )   Save

    BACKGROUND: Studies have shown that extracorporeal shock wave therapy is an effective, safe, and non-invasive treatment for ischemic heart disease, which can improve angiogenesis in the ischemic myocardium.
    OBJECTIVE: To summarize the research advances in promotion of angiogenesis for ischemic myocardium by extracorporeal shock wave therapy.
    METHODS: A computer-based online search of PubMed database and CNKI database was performed for relevant articles published between 1998 and 2014 with key words of “shock wave, ischemic heart disease, angiogenesis, cytokine, stem cell” in English and Chinese, respectively. Articles related to the promotion of angiogenesis for ischemic cardiovascular disease by extracorporeal shock wave were selected. Repetitive articles were excluded. According to inclusion criteria, 51 literatures were selected in result analysis.
    RESULTS AND CONCLUSION: Extracorporeal shock wave therapy can improve angiogenesis in the ischemic myocardium by mobilizing proliferation and differentiation of stem cells into vascular endothelial cells, and by enhancing the expression of growth factors such as vascular endothelial growth factor and basic fibroblast growth factor. Moreover, the extracorporeal shock wave therapy can create a local favorable microenvironment for angiogenesis by inhibiting inflammation, oxidative stress and apoptosis and by regulating components of the extracellular matrix. Extracorporeal shock wave therapy plays an important role in the angiogenesis of ischemic myocardium and displays a good clinical prospect in the treatment of ischemic heart disease. However, the specific mechanism requires further studies.
    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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