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04 March 2016, Volume 20 Issue 10 Previous Issue    Next Issue

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Adipose-derived mesenchymal stem cells regulate T cell immune status in allergic rhinitis Xiao Er-bin, Zhao Bao-jian, Zhang Chi 2016, 20 (10):  1373-1381.  doi: 10.3969/j.issn.2095-4344.2016.10.001 Abstract ( 258 )   PDF (533KB) ( 610 )   Save BACKGROUND: Adipose-derived mesenchymal stem cells are a kind of pluripotent stem cells that have the potential of self-renewal and proliferation, and have low immunogenicity and immunomodulatory role. OBJECTIVE: To study the effects of adipose-derived mesenchymal stem cells on T cell immune status of allergic rhinitis mouse models. METHODS: Sixty mice were randomly assigned into six groups (sensitized/challenged/treatment): experimental group 1 was given ovalbumin/ovalbumin/high-dose adipose-derived mesenchymal stem cells, experimental group 2 given ovalbumin/ovalbumin/low-dose adipose-derived mesenchymal stem cells, experimental group 3 given ovalbumin/ovalbumin/PBS, experimental group 4 given ovalbumin/ovalbumin/0, and experimental group 5 given PBS/PBS/0, and normal control group given no treatment. In the former five groups, intraperitoneal injection of 200 μL ovalbumin sensitizing solution or PBS was conducted for basic sensitization at days 0, 7, 14; 20 μL ovalbumin challenging solution or PBS was given for challenging at days 15-19. In the former three groups, 0.1 mL of high-dose, low-dose adipose-derived mesenchymal stem cells or PBS was given via the tail vein, respectively, at days 20-22 after sensitization and challenge. At 48 hours after final treatment, ELISA was used to detect serum levels of interleukin-4, interleukin-6, interleukin-10 and interferon-γ, and fluorogenic quantitative PCR used to detect the mRNA expressions of these cytokines in the spleen. Migration of fluorescent-labeled adipose-derived mesenchymal stem cells in the nasal mucosa was observed under fluorescence microscope, and pathological changes of the nasal mucosa were observed through hematoxylin-eosin staining. RESULTS AND CONCLUSION: Compared with the experimental group 4, the levels of interleukin-4 and interleukin-6 in the serum and spleen were significantly lower in the experimental group 1 (P < 0.05), and the levels of interluekin-10 and interferon-γ levels were significantly increased (P <0.05); while in the experimental group, the levels of interleukin-6 were reduced significantly (P < 0.05), the levels of interleukin-10 was increased significantly (P < 0.05), but no changes were found in the levels of interleukin-4 and interferon-γ (P > 0.05). Fluorescent-labeled adipose-derived mesenchymal stem cells could migrate into the nasal mucosa, and the number of migrated cells was notably higher in the experimental group 1 than experimental group 2. Eosinophil infiltration in the nasal mucosa was remarkably alleviated in the experimental groups 1 and 2. These findings suggest that adipose-derived mesenchymal stem cells play a non-specific immunomodulatory effect dose-dependently by regulating Th1/Th2 immune imbalances and deficiencies of Treg 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 Figures and Tables | References | Related Articles | Metrics

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Surface labeling of bone marrow mesenchymal stem cells by biotin-streptavidin Yang Lin, Luo Fu-li, Li Yun, Wen Jun, Xu Yang 2016, 20 (10):  1382-1388.  doi: 10.3969/j.issn.2095-4344.2016.10.002 Abstract ( 238 )   PDF (624KB) ( 549 )   Save BACKGROUND: Currently, there is a lack of efficient, non-invasive way to transplant stem cells to the target organ or tissue. Exploring a way to guide targeting transplantation of stem cells and to improve the efficiency of stem cell homing is now one of focuses in the field of stem cells research. OBJECTIVE: To establish a simple and feasible method to chemically modify the cell surface using biotin-streptavidin reaction system, and to evaluate the efficiency of this method to label bone marrow mesenchymal stem cells (BMSCs) and its effects on cell biological functions. METHODS: Passage 3 BMSCs were obtained by whole bone marrow culture method and verified by flow cytometry. Biotin, streptavidin, sulfonated biotin-N-hydroxy-succinimide were used to equip the adhesion molecule ligand, sialyated LewisX (SLeX), to the BMSCs surface. The labeling rate of BMSCs was assessed using fluorescence microscope, the vitality of BMSCs was evaluated by trypan blue staining, and the proliferation of BMSCs was evaluated by cell counting kit-8 assay. Adipogenic and osteogenic inductions were used to evaluate the effect of the method on the multi-differentiation function of BMSCs. RESULTS AND CONCLUSION: After culture for 2 weeks, passage 3 BMSCs were obtained and confirmed by expressing CD90, CD29 and lack of CD34, CD45. Biotin, streptavidin, sulfonated biotin-N-hydroxy-succinimide were successfully used to equip sialyated LewisX (SLeX) to the BMSCs surface and had minor effects on the vitality, proliferation, and differentiation of BMSCs. This method was simple for surface modification and had a high modification rate of 88%. The homing of BMSCs modified by this method to the target organ or tissue could be greatly enhanced. Therefore, this method potentially could have extensive and important applications.   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  Figures and Tables | References | Related Articles | Metrics

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Activation of Wnt signaling pathway in the neuronal differentiation of bone marrow mesenchymal stem cells: a microarray analysis Lin Zhi-ping, Zeng Rong, Lin Hao 2016, 20 (10):  1389-1395.  doi: 10.3969/j.issn.2095-4344.2016.10.003 Abstract ( 309 )   PDF (555KB) ( 606 )   Save BACKGROUND: It will provide a new insight into the future application of bone marrow mesenchymal stem cells in the treatment of spinal cord injury and tissue engineering by studying the effect of activation of Wnt signaling pathway in the neuronal differentiation of bone marrow mesenchymal stem cells. OBJECTIVE: To detect the expression of related genes by gene chip technology during the neuronal differentiation of bone marrow mesenchymal stem cells. METHODS: Human bone marrow mesenchymal stem cells were isolated and purified, and passage 5 cells were obtained. GatewayTM technology was used to build lentiviral vectors that was used to transfect Wnt-1 into human bone marrow mesenchymal stem cells. Control, non-transduction and transduction groups were set in this study. Human bone marrow mesenchymal stem cells were then induced to differentiate into neurons. Cell morphology was observed under inverted phase contrast microscope. Gene chip was used to detect the regulation changes and the differential expression of related genes in the Wnt signaling pathway. RESULTS AND CONCLUSION: Under the scanning electron microscope, the transfected cells were found to have the similar morphology of neuron-like cells. Analysis by the gene chip hybridization technique showed that 3 287 genes were up-regulated and 4 215 genes were down-regulated in the signal pathway. In the Wnt signaling pathway, genes related to the nervous system development and differentiation were up- or down-regulated. It is verified that the Wnt signal pathway is activated via Wnt-1 transduction, and the downstream genes appear to have genetic transcription so as to promote the neuronal differentiation of human 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 Figures and Tables | References | Related Articles | Metrics

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Effects of astragalus polysaccharides on the proliferation of bone marrow mesenchymal stem cells from patients with multiple myeloma Yang Jing-ke, Lv Feng-shou, Han Li 2016, 20 (10):  1396-1401.  doi: 10.3969/j.issn.2095-4344.2016.10.004 Abstract ( 356 )   PDF (473KB) ( 371 )   Save BACKGROUND: Bone marrow mesenchymal stem cells can produce a variety of cytokines to provide some support for the growth and proliferation of myeloma cells. OBJECTIVE: To investigate the effects of astragalus polysaccharides on the proliferation of bone marrow mesenchymal stem cells from patients with multiple myeloma and cytokine secretion. METHODS: Bone marrow mesenchymal stem cells were isolated by density gradient centrifugation from the bone marrow of a healthy volunteer and a multiple myeloma patient, and were cultured and identified. Passage 3 bone marrow mesenchymal stem cells were collected, and the growth curve was depicted by the enzyme marker method. MTT method and flow cytometry were used to detect the effects of different concentrations (0.5, 1, 2, 4, 6, 8 g/L) of astragalus polysaccharides on the proliferation and cell cycle of bone marrow mesenchymal stem cells. The expression levels of interleukin-1β and interleukin-6 were detected by ELISA method. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells from both the healthy control and multiple myeloma patient showed negative expression of CD34 and positive expression of CD44. The obtained cells from two sources had the characteristics of bone marrow mesenchymal stem cells. Cells from the healthy control had significantly shorter doubling time than those from the myeloma patient. After treated with 1 and 2 g/L astragalus polysaccharide, cell proliferation rate in the multiple myeloma group was significantly higher than that in the healthy control group; after treated with 6 and 8 g/L astragalus polysaccharide, the cell proliferation rate was significantly lower in the multiple myeloma group than in the healthy control group. After treated with 1 g/L astragalus polysaccharide, compared with the healthy control, the proportion of bone marrow mesenchymal stem cells from the multiple myeloma patient in G0/G1 phase was significantly decreased, while the proportions of cells in both S phase and G2/M phase from the myeloma patient were significantly increased. Interleukin-1β and interleukin-6 levels of bone marrow mesenchymal stem cells from the patient with multiple myeloma were significantly higher than those of cells from healthy control. After intervention with 0.5, 1, 2 g/L astragalus polysaccharide, the interleukin-1β and interleukin-6 levels of bone marrow mesenchymal stem cells from the patient with multiple myeloma were all significantly decreased. In conclusion, a certain concentration of astragalus polysaccharides can promote the proliferation of bone marrow mesenchymal stem cells and decrease the levels of interleukin-6 and interleukin-β in patients with multiple myeloma.  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 Figures and Tables | References | Related Articles | Metrics

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Stromal cell derived factor-1 promotes the proliferation of bone marrow stem cells: the optimal concentration is 200 μg/L Peng Wei, Qin Yuan, Liao Chun-hui, Chen Song-ling 2016, 20 (10):  1402-1408.  doi: 10.3969/j.issn.2095-4344.2016.10.005 Abstract ( 265 )   PDF (665KB) ( 248 )   Save BACKGROUND: Stromal cell derived factor-1 is a small molecular protein with a wide range of biological activity that can cause immune cell chemotaxis, and it also has a chemotactic effect on bone marrow stem cells and periodontal ligament cells. OBJECTIVE: To investigate the effect of stromal cell derived factor-1 with different concentrations on the proliferation of bone marrow stem cells and to probe the best concentration. METHODS: Bone marrow stem cells from beagle dogs were cultured in vitro and stimulated by different concentrations of stromal cell derived factor-1 (100, 200, 300 μg/L). MTT was used to detect the influence of stromal cell derived factor-1 on the proliferation of bone marrow stem cells so as to screen the best concentration of stromal cell derived factor-1. Then, stromal cell derived factor-1 at the best concentrations was used to intervene the bone marrow stem cells, and MTT was used again to detect the proliferation of bone marrow stem cells. RESULTS AND CONCLUSION: Stromal cell derived factor-1 at concentrations of 100, 200, 300 μg/L could promote the proliferation of bone marrow stem cells, and the effect was more notable at 200 and 300 μg/L but with no significant difference. Therefore, 200 μg/L was considered to be the best concentration of stromal cell derived factor-1 for intervention of bone marrow stem cells. Compared with the blank control group, 200 μg/L stromal cell derived factor-1 could significantly promote the proliferation of bone marrow stem cells. Taken together, stromal cell derived factor-1 can promote the proliferation of bone marrow stem cells, and its best concentration is 200 μg/L.  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 Figures and Tables | References | Related Articles | Metrics

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Overexpression of trophoblastic stem cell transcription factor, forkhead box D3, contributes to malignancy of human choriocarcinoma JAR cells Liu Yuan, Wang Ya-xin, Wu Wei-bin, Wang Yu-dong, Zhang Hui-juan 2016, 20 (10):  1409-1418.  doi: 10.3969/j.issn.2095-4344.2016.10.006 Abstract ( 188 )   PDF (667KB) ( 263 )   Save BACKGROUND: Choriocarcinoma is a kind of trophoblastic neoplasm with highly aggressive phenotypes. Forkhead box D3 (FoxD3) is an embryonic and trophoblastic stem cell transcription factor. It plays important roles in different physical and pathological situations such as embryogenesis, carcinogenesis and tumor progression. OBJECTIVE: To investigate the role of FoxD3 in choriocarcinoma malignancy and the possible mechanism. METHODS: The human choriocarcinoma JAR cell line was employed in this study. The mRNA and protein expressions of genes were measured by quantitative RT-PCR (qRT-PCR) and western blot, respectively. The FoxD3 specific short hair RNA was applied to down-regulate gene expression. The cell proliferation was evaluated in vitro by cell counting assay and in vivo by tumor growth. The migration/invasion was determined by transwell assay. The profile of FoxD3 targeted genes was investigated with an Agilent microarray and verified by qRT-PCR. RESULTS AND CONCLUSION: The FoxD3 mRNA and protein expressions in JAR cells were significantly higher than those in primarily cultured normal trophoblastic cells. Knockdown of FoxD3 by short hair RNA in JAR cells could inhibit cell proliferation and migration/invasion in vitro, and suppress the tumor growth with decreased β-human chorionic gonadotropin secretion in vivo. A profile of seven focal adhesion molecules (ITGA5, ITGB6, THBS4, COL6A3, VTN, NRXN3 and NLGN1) was verified to be targeted by FoxD3. Furthermore, knockdown of FoxD3 by short hair RNA could decrease the activation of focal adhesion kinase. All these findings suggest the overexpression of FoxD3 can contribute to the aggressive phenotype of choriocarcinoma JAR cells by regulating the profile of focal adhesion molecules and focal adhesion kinase.    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 Figures and Tables | References | Related Articles | Metrics

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Polylysine-modified gamma-Fe2O3 nanoparticle labeling has no effect on neuroblastoma stem cell activation and proliferation Zhong Zhi-yong, Shi Bao-jun, Zhou Hui, Wang Wen-bo 2016, 20 (10):  1419-1425.  doi: 10.3969/j.issn.2095-4344.2016.10.007 Abstract ( 255 )   PDF (561KB) ( 673 )   Save BACKGROUND: Retinoic acid is the most promising inducer for neuroblastoma minimal residual lesion, and it can induce cell differentiation in vivo, accompanied by reducing tumor cell proliferation. OBJECTIVE: To study the effect of nanoparticle labeling on biological characteristics of neuroblastoma stem cells, and the role of 13-cis retinoic acid to induce differentiation of neuroblastoma stem cells. METHODS: Neuroblastoma stem cells were isolated and cultured in vitro using serum-free suspension culture method, labeled with polylysine-modified γ-Fe2O3 nanoparticles and induced in culture medium containing 13-cis retinoic acid. RT-PCR was used to detect the expression of Oct-4 before and after labeling as well as before and after induction. Immunofluorescence method was used to detect the expression of nestin before and after labeling as well as before and after induction. RESULTS AND CONCLUSION: Neuroblastoma stem cells were successfully cultured in the bone marrow samples from 5 of 20 cases. Polylysine-modified γ-Fe2O3 nanoparticle labeling did no influence the viability and proliferation ability of neuroblastoma stem cells, and also had no effect on Oct-4 mRNA and nestin expression. After cultured in the culture medium containing 13-cis retinoic acid, the cell shape changed and the growth rate slowed down. Moreover, the expression of Oct-4 mRNA and nestin was gradually reduced. These findings indicate that polylysine-modified gamma-Fe2O3 nanoparticles can be used to label neuroblastoma stem cells, and 13-cis retinoic acid can induce the differentiation of neuroblastoma 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 Figures and Tables | References | Related Articles | Metrics

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Role of epithelial-mesenchymal transition in transformation of gastric cancer cells SGC7901 to gastric cancer stem cells Xing Hui-jun, Zhao Yan-jun, Hou Lei, Sun Yong, Liu Peng, Li Chun-hui 2016, 20 (10):  1426-1432.  doi: 10.3969/j.issn.2095-4344.2016.10.008 Abstract ( 215 )   PDF (497KB) ( 487 )   Save BACKGROUND: Studies have found that epithelial-mesenchymal transition is closely related with tumor invasion, metastasis, and drug resistance, but studies on the role of epithelial-mesenchymal transition in the transformation process of gastric cancer cells SGC7901 to gastric cancer stem-like cells are rarely reported. OBJECTIVE: To explore the effect of epithelial-mesenchymal transition in the transformation process of gastric cancer cells SGC7901 to gastric cancer stem-like cells. METHODS: Vincristine-induced SGC7901 cells were cultured and screened to prepare gastric cancer stem-like cells. CD44 phenotype, morphological changes, stem cell-related markers, and epithelial-mesenchymal transition related molecules were detected. RESULTS AND CONCLUSION: After passage, vincristine-induced SGC7901 cells presented with morphological changes, and clonal cell spheres generated after serum-free suspension culture. Meanwhile, the proportion of SGC7901 cells positive for CD44 was decreased. Expression levels of SOX2, OCT4, Snail1 mRNA, Twist mRNA and Vimentin mRNA were significantly higher in the gastric cancer stem-like cells than SGC7901 cells, but the expression level of E-caderin was lower in the gastric cancer stem-like cells than SGC7901 cells. These findings indicate that gastric cancer cells SGC7901 can be successfully transformed into gastric cancer stem-like cells, and the epithelial-mesenchymal transition is involved in this transforming progress.   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   Figures and Tables | References | Related Articles | Metrics

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Cryopreservation of bone marrow mesenchymal stem cells from Sprague-Dawley rats at -80 ℃ Chen Kai, Li Xin-ran, Xuan Yan 2016, 20 (10):  1433-1438.  doi: 10.3969/j.issn.2095-4344.2016.10.009 Abstract ( 303 )   PDF (475KB) ( 306 )   Save BACKGROUND: We attempt to explore a low-cost, simple and effective way to cryopreserve bone marrow mesenchymal stem cells at -80 ℃. OBJECTIVE: To screen the optimal cryopreservation fluid for bone marrow mesenchymal stem cells and to verify the biological features of bone marrow mesenchymal stem cells after long-term cryopreservation. METHODS: Bone marrow mesenchymal stem cells were cultured using adherent method and the biological features and purity of cells were detected using immunofluorescence method. Bone marrow mesenchymal stem cells were cryopreserved in the cryoprotectant medium containing low-sugar DMEM, fetal bovine serum and dimethyl sulfoxide at different proportions at -80 ℃ for a short term. Then, the optimal cryoprotectant was selected to storage the bone marrow mesenchymal stem cells. After 1, 3, 6 months of cryopreservation, the cells were resuscitated, cultured and passaged. Passage cells were identified immunofluorescence method to determine the biological features of bone marrow mesenchymal stem cells cryopreserved at -80 ℃. RESULTS AND CONCLUSION: Cryoprotectant medium of 80% DMEM+10% fetal bovine serum+10% dimethyl sulfoxide was suitable for cryopreserving MSCs at -80 ℃, and resuscitated cells were able to proliferate in vitro, and passage normally, indicating the cryopreserved bone marrow mesenchymal stem cells still maintain the original biological activity.  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 Figures and Tables | References | Related Articles | Metrics

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The homing of transplanted bone marrow mesenchymal stem cells to the thymus Wang Zhi-hong, Chen Wei-min, Guo Kun-yuan 2016, 20 (10):  1439-1445.  doi: 10.3969/j.issn.2095-4344.2016.10.010 Abstract ( 309 )   PDF (661KB) ( 409 )   Save BACKGROUND: Bone marrow mesenchymal stem cells have low immunogenicity and can induce immune tolerance. At present, the mechanism of immune regulation of bone marrow mesenchymal stem cells is not completely understood. It has been rarely reported whether the bone marrow mesenchymal stem cells can migrate to the thymus after transplantation. OBJECTIVE: To observe the distribution and survival of bone marrow mesenchymal stem cells in the thymus of aging rats after transplantation. METHODS: Bone marrow mesenchymal stem cells cultured in vitro were transfected by adenovirus vectors expressing green fluorescent protein. Transfected bone marrow mesenchymal stem cells were injected into the portal vein of aging rats. At days 3, 7, 14, 21 after transplantation, the survival of bone marrow mesenchymal stem cells homing to the thymus was observed under fluorescence microscope. At day 3 after transplantation, thymus tissues were taken and stained with hematoxylin-eosin for pathological observation. RESULTS AND CONCLUSION:Green fluorescent protein-labeled bone marrow mesenchymal stem cells had a strong green fluorescence at days 3 and 7 after transplantation, and the cell contour was clear. There was no significant difference in the mean absorbance values at days 3 and 7 (P > 0.05). Expression of green fluorescent protein was weakened significantly at days 14 and 21 compared with that at day 3 (P < 0.05). At 3 days after transplantation, the transplanted bone marrow mesenchymal stem cells were clearly visible in the thymus, and acute rejection was not observed. The results show that bone marrow mesenchymal stem cells can migrate to the damaged thymus tissue through the blood circulation, and can survive at least 1 week.  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 Figures and Tables | References | Related Articles | Metrics

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Stereotactic transplantation of neural stem cells into the brain improves motor function of craniocerebral trauma rats Diao Xin-feng, Cheng Li-min, Xue Yong, Hu Cheng-wang, Cai Zhong-li 2016, 20 (10):  1446-1451.  doi: 10.3969/j.issn.2095-4344.2016.10.011 Abstract ( 371 )   PDF (515KB) ( 373 )   Save BACKGROUND: Cell replacement therapy as an effective strategy for reconstruction of the central nervous system has very broad application prospects. OBJECTIVE: To investigate the effect of stereotactic transplantation of neural stem cells into the brain on the neuromotor function of craniocerebral trauma rats. METHODS: Twenty male Sprague-Dawley rats were equivalently randomized into study and control groups. Animal models of craniocerebral trauma were made using the improved free-fall method in the rats. Then, model rats in the study and control groups were given parenchymal transplantation of embryonic neural stem cells and the same volume of culture medium with no stem cells at 1 day after injury, respectively. Neuromotor function of rats was assessed based on the neurological severity scores. At 2 weeks after transplantation, brain tissues were taken for hematoxylin-eosin staining, anti-BrdU, glial fibrillary acidic protein, β-tubulin III and tyrosine hydroxylase immunohistochemistry staining. RESULTS AND CONCLUSION: The neurological severity scores in the study group were significantly lower than those in the control group at 1 and 2 weeks after injury (P < 0.05). In the study group, there were many BrdU-positive neural stem cells in the brain tissues, some of which were positive for glial fibrillary acidic protein, β-tubulin III and tyrosine hydroxylase; while in the control group, there was no BrdU-positive cell in the brain tissues. Experimental findings show that neural stem cells stereotactically transplanted into the brain can proliferate and differentiate in the brain lesion, and thereby notably improve the neuromotor function of rats with craniocerebral trauma.  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   Figures and Tables | References | Related Articles | Metrics

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Pretreatments with hypertonic solution and cobalt chloride in bone marrow mesenchymal stem cell transplantation for treatment of degenerative disc disease Ye Han, Meng Zhen, Lin Jia-chen, Li Jia-wei, Zhang Yong-xing, Lin Nan-he, Zhao Qing-hua 2016, 20 (10):  1452-1460.  doi: 10.3969/j.issn.2095-4344.2016.10.012 Abstract ( 266 )   PDF (658KB) ( 461 )   Save BACKGROUND: Stem cell therapy has been used for prevention and treatment of degenerative disc disease. Considering the special microenvironment in the intervertebral disc, the survival rate and differentiation ability of transplanted cells are decreased, which may lead to the poor efficacy of stem cell therapy. How to improve the survival ability and therapeutic effect of the transplanted cells is the focus of stem cell therapy for degenerative disc disease. OBJECTIVE: To investigate the effects of cobalt chloride combined with hypertonic solution pretreatment on bone marrow mesenchymal stem cells that will be transplanted for treatment of degenerative disc disease. METHODS: (1) In vitro cell experiment: bone marrow mesenchymal stem cells were divided into three groups and subjected to normal culture medium (normal control group), 1% hypertonic mother solution (hypertonic group), 100 μmol/L cobalt chloride (hypoxia group), or 1% hypertonic mother solution plus 100 μmol/L cobalt chloride (combined group) for 1 week. Then, 2% hypertonic solution and 200 μmol/L cobalt chloride cobalt chloride were used to simulate the anaerobic and hypertonic environment intervenes in pretreated and untreated bone marrow mesenchymal stem cells for 24 hours. After that, RT-PCR was used to detect the expression of caspase-3 for apoptosis evaluation. (2) In vivo animal experiment: Sprague-Dawley rats were divided into model, cell transplantation and hypertonic plus hypoxic groups. Rat models of intervertebral disc degeneration were made in these three groups. After modeling, rats in these three groups were given no treatment, bone marrow mesenchymal stem cell transplantation or transplantation of bone marrow mesenchymal stem cells which were subjected to hypertonic and hypoxia pretreatments into the intervertebral disc. Two weeks later, immunohistochemistry and RT-PCR methods were used to detect cell distribution and related gene expression, respectively, thereby to evaluate the therapeutic effect of stem cells. RESULTS AND CONCLUSION: (1) In vitro cell experiment: caspase-3 mRNA expression was significantly reduced in pretreated bone marrow mesenchymal stem cells compared with the untreated cells (P < 0.05). (2) In vivo animal experiment: compared with the control group, the caspase-3 and interleukin-1β in the intervertebral disc and a number of degenerative indexes were decreased in the cell transplantation. Compared with the cell transplantation group, these indicators had better outcomes in the hypertonic plus hypoxic group (P < 0.05). These findings indicate that bone marrow mesenchymal stem cells have therapeutic potential for degenerative disc disease, and have better adaptability and transplantation effects by hypertonic and hypoxia pretreatments.  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 Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cell transplantation for the treatment of systemic lupus erythematosus Wang Zhi-guo, Liu Xue-ming, Tong Sheng-quan, Shi Zhe-qun, Rao Li 2016, 20 (10):  1461-1467.  doi: 10.3969/j.issn.2095-4344.2016.10.013 Abstract ( 310 )   PDF (550KB) ( 542 )   Save BACKGROUND: The non-specific immune suppression method is generally used for treatment of systemic lupus erythematosus, but poor prognosis, such as infection and high recurrence rate, exists. OBJECTIVE: To evaluate the therapeutic effect of bone marrow mesenchymal stem cell transplantation on systemic lupus erythematosus in mice. METHODS: Sixteen mice with systemic lupus erythematosus were equivalently randomized into control and experimental groups, or then subjected to passage 3 bone marrow mesenchymal stem cell transplantation or the equal volume of normal saline via the tail vein, respectively. Mouse urine samples were collected to detect urine protein levels by Bradford method. Blood samples from the tip of the mouse tail were extracted to detect serum anti-ds-DNS antibody concentration by radioimmunoassay. Mouse kidney tissues were taken and observed pathohistologically through hematoxylin-eosin staining and immunohistochemistry staining under microscope. Flow cytometry was used to detect the expression of CD4+CD25+T cells in the inner canthus blood, fresh spleen and thymus. RESULTS AND CONCLUSION: Within 10 weeks after cell transplantation, the urine protein levels in the two groups were gradually increased, and the rising velocity was higher in the control group than in the experimental group. From the 4th to 10th week, the urine protein levels in the experimental group were significantly lower than those in the control group (P < 0.05). In the control group, lymphocyte infiltration was visible in the kidney tissues with a few of plasmocytes, and pathological findings showed the mice presented with interstitial nephritis; in the experimental group, the mice had no pathological changes in the kidney. In the two groups, immune complexes were found in the mesangial area, which showed a patch-like distribution in the control group and a punctate distribution in the experimental group; the relative proportion of the occupied area in the experimental group was significantly lower than that in the control group. The expression level of CD4+CD25+T cells in the blood and thymus were significantly higher in the experimental group than the control group (P < 0.05), and the expression level of CD4+CD25+T cells in the spleen was slightly higher in the experimental group than the control group with no significant difference (P > 0.05). The serum anti-ds-DNA antibody concentration in the experimental group was significantly lower than that in the control group (P < 0.05). Taken together, bone marrow mesenchymal stem cell transplantation can improve the pathological damage in systemic lupus erythematosus mice, and has a certain therapeutic effect on systemic lupus erythematosus.   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 Figures and Tables | References | Related Articles | Metrics

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Biological features of CD90+ tumor stem cells in ovarian cancer Liu Zhi-hui, Xiang Zhi-hui 2016, 20 (10):  1468-1473.  doi: 10.3969/j.issn.2095-4344.2016.10.014 Abstract ( 307 )   PDF (480KB) ( 384 )   Save BACKGROUND: Thinking from ovarian cancer stem cell theory shows that: in the tumor cells, there are a fraction of stem cells with self-renewing ability and multipotent differentiation, which are the root causes of ovarian cancer recurrence and drug resistance. Studies have shown that CD90 can be used as a surface marker of mesenchymal stem cells and stem cells of other cancers. OBJECTIVE: To explore the biological features of CD90+ tumor cells from ovarian cancer tissues. METHODS: Primary ovarian cancer cells were isolated from the abdominal dropsy of ovarian cancer patients to sort CD90+ and CD90- cells using flow cytometry. RT-PCR was used to detect expressions of stem cell-related genes and epithelial to mesenchymal transition-related genes. Cell invasion was observed by Transwell invasion assay, cell proliferation and differentiation observed by clone formation assay, stem cell potential observed by suspension sphere-forming assay, and tumor formation rate observed by in vivo tumorigenicity experiment. RESULTS AND CONCLUSION: Compared with the CD90- cells, the expressions of CD44, CD133, ALDH1, N-cad and Vimentine were significantly higher in the CD90+ cells (P < 0.05), but the expression of E-cad was significantly decreased in the CD90+ cells (P < 0.05). Tumor formation rates of CD90- and CD90+ cells were increased significantly with the increase of seeded cell number, which was more obvious in CD90+ cells. The number of transmembrane cells, the number of cell clones and the number of suspended spheres were significantly higher in the CD90+ cells than the CD90- cells (P < 0.05). Experimental findings from this study show that CD90+ cells highly express epithelial to mesenchymal transition-related genes and stem cell-related genes, with higher invasion, proliferation and differentiation, in vivo tumorigenicity and potential of stem cells. CD90+ cell separation may be a new method to separate ovarian cancer 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 Figures and Tables | References | Related Articles | Metrics

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Neural stem cell transplantation for sequela of traumatic brain injury: the best timing for treatment Lou Yong-li, Chen Ping, Jiang Yu, Zhang Hui, Min You-hui 2016, 20 (10):  1474-1480.  doi: 10.3969/j.issn.2095-4344.2016.10.015 Abstract ( 347 )   PDF (536KB) ( 549 )   Save BACKGROUND: Neural stem cell transplantation provides an important way to treat sequela of traumatic brain injury, but the timing for treatment is inconclusive. OBJECTIVE: To explore the clinical effect of neural stem cell transplantation in the treatment of sequela of traumatic brain injury and the choice of the best treatment time. METHODS: Totally 178 patients with sequela of traumatic brain injury who underwent neural stem cell transplantation were divided into three groups as per the timing for neural stem cell transplantation: group A (with 6 months after injury, n=60), group B (6-12 months after injury, n=59), and group C (over 12 months after injury, n=59). Improvement in clinical symptoms and scores on function independent measure (FIM) were recorded and compared in the three groups. RESULTS AND CONCLUSION: The total effective rate of group A was significantly higher than that in groups B and C (P < 0.05). FIM scores were significantly improved in the three groups after cell transplantation (P < 0.05). At 3 months after the fourth transplantation, the FIM score in the group A was significantly higher than that in the other two groups, and the incidence of adverse reactions in the group A was significantly lower than that in the other two groups (P < 0.05). These findings indicate that neural stem cell transplantation at different timing can all harvest certain clinical effects, but the best timing for neural stem cell transplantation is within 6 months after injury.  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 Figures and Tables | References | Related Articles | Metrics

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Hydrogen peroxide accelerates senescence of human dental pulp stem cells Xu Ke, Feng Gui-juan, Feng Xing-mei, Huang Dan, Zheng Ke, Tang En-yi 2016, 20 (10):  1481-1487.  doi: 10.3969/j.issn.2095-4344.2016.10.016 Abstract ( 407 )   PDF (558KB) ( 458 )   Save BACKGROUND: The process of oxidative stress that impacts the curative effect exists in the region which accepts cell transplantation. However, there are few reports about the effects of oxidative stress on human dental pulp stem cells and relevant mechanism. OBJECTIVE: To understand the effect of hydrogen peroxide on the senescence of human dental pulp stem cells. METHODS: Human dental pulp stem cells were isolated and cultured in PBS, 100 and 200 μmol/L hydrogen peroxide for 2 hours, respectively. Cell morphology was observed under inverted microscope, degree of cell senescence monitored by β-galactosidase staining, cell proliferation ability detected by BrdU kit and cell counting method, cytoskeleton of dental pulp stem cells and expression of sirt1 tested using immunofluorescence method, and expression of sirt1 and p16 proteins measured by western blot assay. RESULTS AND CONCLUSION: Dental pulp stem cells exhibited a fibroblast-like morphology with spindle-shaped appearance. After stimulated by hydrogen peroxide, the cell volume was enlarged, the β-galactosidase staining deepened and the proliferation of dental pulp stem cells reduced. The enhancement of senescence of dental pulp stem cells was accompanied with the increasing concentration of hydrogen peroxide, and in this process, the expression of p16 was raised while the expression of sirt1 was decreased. In conclusion, the senescence of human dental pulp stem cells can be promoted by the stimulation of hydrogen peroxide, and sirt1 and p16 are involved in this process. Our findings may provide a theoretical and experimental foundation for autologous transplantation of dental pulp 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 Figures and Tables | References | Related Articles | Metrics

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Isoflurane inhibits neural stem cell proliferation in the hippocampus and promotes its differentiation into neurons Sun Nai, Li Chun-wei, Zhao Wei-xin, Song Qiong, Xia Gui-shan 2016, 20 (10):  1488-1493.  doi: 10.3969/j.issn.2095-4344.2016.10.017 Abstract ( 329 )   PDF (556KB) ( 242 )   Save BACKGROUND: Isoflurane cannot only induce a wide range of large neuronal apoptosis, but also inhibit hippocampal neurogenesis in neonatal rats, thereby resulting in hippocampus-dependent learning and memory defects. OBJECTIVE: To investigate the isoflurane effect on proliferation and differentiation of the hippocampal neural stem cells. METHODS: Twenty-six Sprague-Dawley rats were randomly divided into air group and isoflurane group (n=13 per group). Rats in the isoflurane group were subjected to 2.5% isoflurane inhalation for 3 minutes followed by 1.5% isoflurane inhalation for 4 hours. Rats in the air group only breathed in air. After the intervention, blood glucose and arterial blood gas changes were detected in the two groups. Additionally, rats in the two groups were given intraperitoneal injection of 5-bromodeoxyuridine before and after intervention. At 24 hours after the last injection of 5-bromodeoxyuridine, brain tissues were taken to make frozen sections for immunofluorescence staining. RESULTS AND CONCLUSION: There were no significant difference in pH, PaO2, PaCO2, HCO3, BE and SaO2 levels between the two groups (P > 0.05). Compared with the air group, the number of BrdU+ cells was significantly less in the isoflurane group (P < 0.05), while the number of NeuroD+/BrdU+ cells was significantly higher in the isoflurane group (P < 0.05). The incidence of adverse reactions was 23% in the isoflurane group, which was significantly higher than that in the air group (7.7%; P < 0.05). These findings indicate that isoflurane can inhibit the proliferation of neural stem cells in the hippocampal dentate gyrus, and promote their differentiation 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 Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cell transplantation improves airway inflammation due to chronic asthma Zhang Qi, Guo Rui-rui, Hu Jiang-ping 2016, 20 (10):  1494-1500.  doi: 10.3969/j.issn.2095-4344.2016.10.018 Abstract ( 210 )   PDF (633KB) ( 334 )   Save BACKGROUND: Studies have shown that bone marrow mesenchymal stem cell transplantation can improve disease conditions by reducing inflammation. OBJECTIVE: To explore the therapeutic efficacy of bone marrow mesenchymal stem cells on chronic asthma rats. METHODS: A rat model of chronic asthma was established by intraperitoneally injected and aerosolized ovalbumin. After modeling, rats were given 4×105 and 8×105 bone marrow mesenchymal stem cells via the tail vein, respectively. Thirty days later, the lung tissues were observed pathologically using hematoxylin-eosin staining; RT-qPCR and ELISA methods were employed to test the changes in interleukin-10, tumor necrosis factor-α and interferon-γ levels in lung tissue and peripheral blood, respectively. RESULTS AND CONCLUSION: Rat models of chronic asthma were successfully established after intraperitoneal injection of ovalbumin combined with aerosolized ovalbumin. After 30 days of cell treatment, the structure of lung tissues were obviously recovered, and the levels of interleukin-10, tumor necrosis factor-α and interferon-γ showed some improvement in lung tissue and peripheral blood, but there were no differences between the two groups. In conclusion, bone marrow mesenchymal stem cells show some potential role in the treatment of chronic asthma.    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 Figures and Tables | References | Related Articles | Metrics

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Rhodiola polysaccharide effect on spermatogonial stem cell proliferation in vitro Li Jun-tao, Zhang Pei-hai, Qu Xiao-wei, Li Zheng-sheng, Li Song-wei 2016, 20 (10):  1501-1507.  doi: 10.3969/j.issn.2095-4344.2016.10.019 Abstract ( 245 )   PDF (566KB) ( 250 )   Save BACKGROUND: To establish a rapid and effective method to obtain sufficient spermatogonial stem cells that can meet the clinical need is urgent to be solved in the spermatogonial stem cell transplantation. OBJECTIVE: To study the effect of rhodiola polysaccharide on the proliferation of spermatogonial stem cells in vitro. METHODS: Under sterile conditions, spermatogonial stem cells and Sertoli cells were isolated from the testis of mice, and spermatogonial stem cells were seeded onto the feed layer of Sertoli cells. Then, the co-cultured cells were assigned into experimental group 1 (simple cell culture medium), experimental group 2 (cell culture medium containing 150 mg/L rhodiola polysaccharide) and experimental group 3 (cell culture medium containing 150 mg/L rhodiola polysaccharide, 1 U/L leukemia inhibitory factor and 10 μg/L glial cell line-derived neurotrophic factor). After 7 days of co-culture, flow cytometry was used to detect cell proliferation in vitro, and cell viability and positive expression of GFRa-1, Thy-1 and C-kit were calculated. RESULTS AND CONCLUSION: After 7 days of co-culture, the cells grew rapidly and presented with colony and thyrsiform growth, and the number of cell masses increased significantly, all of which were in line with the proliferative features of spermatogonial stem cells. The GFRa-1, Thy-1 and C-kit proteins were expressed in the cell membrane and cytoplasm, mainly in the cell membrane. The viability of spermatogonial stem cells and positive expression of GFRa-1 and Thy-1 were ranked as follows: experimental group 3 > experimental group 2 > experimental group 1, and there were significant differences between groups (P < 0.05). The positive expression of C-kit had no difference between experimental groups 1 and 2, but it was significantly higher in the experimental group 3 than the other two groups (P < 0.05). These findings indicate that rhodiola polysaccharide used alone or combined with leukemia inhibitory factor and glial cell line-derived neurotrophic factor can enhance the proliferative ability of spermatogonial stem cells in vitro.  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 Figures and Tables | References | Related Articles | Metrics

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Isolation, purification and preservation of adipose-derived stem cells: research progress and future development Chen You-bai, Chen Cong-hui, Qixu Zhang, Han Yan 2016, 20 (10):  1508-1520.  doi: 10.3969/j.issn.2095-4344.2016.10.020 Abstract ( 633 )   PDF (865KB) ( 2326 )   Save BACKGROUND: In 2001, Zuk et al found adipose-derived stem cells (ASCs) from the aspirate of liposuction for the first time, which launched a new era of stem cell research. In recent years, stem cells have been proved to widely exist in many tissues and organs. ASCs are always in the spotlight of plastic and reconstructive surgery, tissue engineering and regenerative medicine because of extensive sources and simple isolation. OBJECTIVE: To review the fat tissue harvesting and ASCs isolation, purification, expansion, and cryopreservation, to discuss the main factors which influence the yield, proliferation capacity and differentiation potential of ASCs, and to predict the future research interests based on current issues. METHODS: On September 10th, 2015, relevant articles were searched in PubMed using the following format: (adipose stem cells[Title]) OR (adipose-derived stem cells[Title]) OR (adipose-derived mesenchymal stem cells[Title]) and in SinoMed using the following format in Chinese: (“adipose-derived stem cells” [Title])or(“adipose-derived mesenchymal stem cells”[Title]). Finally, 81 representative articles were included according to their titles and abstracts. In this review, we also introduced relevant experience about the aforementioned procedures from the Department of Plastic Surgery and Tissue Regeneration and Molecular Cell Engineering Lab of University of Texas, MD Anderson Cancer Center, USA. RESULTS AND CONCLUSION: The widely dispersed fat tissues potentially provide abundant stem cells for tissue engineering and regenerative medicine. Liposuction is a mini-invasive approach for harvesting fat tissues. Collagenase digestion is the major method for ASCs isolation due to its simplicity and high yield in basic research. However, clinical fat transplantation without ASCs isolation or non-collagenase isolation of stromal vascular fraction or ASCs is preferred. The phenotype, proliferation and differentiation capacity of ASCs may be affected by several factors during the fat tissue harvesting and ASCs isolation. Therefore, a standard protocol for ASCs isolation is needed.   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 Figures and Tables | References | Related Articles | Metrics