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05 February 2014, Volume 18 Issue 6 Previous Issue    Next Issue

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Recombinant lentiviral vector transfected sheep bone marrow mesenchymal stem cells and osteogenic gene expression changes Han Xiang-zhen, He Hui-yu, Hu Yang, Ba Jiao-jiao, Wang Huan-huan, Mi Xue, Abulizi•Abudula 2014, 18 (6):  821-828.  doi: 10.3969/j.issn.2095-4344.2014.06.001 Abstract ( 314 )   PDF (2437KB) ( 471 )   Save  BACKGROUND:Basic fibroblast growth factor (bFGF) can promote the proliferation of bone marrow stromal cells, and bone morphogenetic protein-2 (BMP-2) has an important significance in the induction of new bone formation. OBJECTIVE: To analyze the effects of bFGF, BMP-2 and their co-transfection on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and to compare the relative expressions of collagen I, osteocalcin and osteopontin before and after cell transfection, thereby providing theoretical implications for seed cells in the construction of tissue-engineered bone. METHODS:Lentiviral vectors carrying bFGF and BMP-2 were constructed to transfect sheep bone marrow mesenchymal stem cells. Cells were divided into four groups: bFGF group, BMP-2 group, co-transfection group and control group. The RNA was extracted using real-time quantitative PCR to detect mRNA levels of collagen I, osteocalcin, and osteopontin. RESULTS AND CONCLUSION: Significant difference in non-specific osteogenic gene expressions was found among the four groups (P < 0.05). bFGF and BMP-2 showed an interaction (P < 0.05). Expressions of collagen I and osteocalcin in the co-tranfection group were higher than those in the other three groups (P < 0.05), but osteopontin expression exhibited no difference (P > 0.05). In vitro experiments showed that the relative expression of collagen I, osteocalcin and osteopontin were higher in the co-transfection group, indicating the cells from the co-transfection group have strongest osteogenic capacity that are suitable for seed cells for 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cells differentiate into neuron-like cells induced by combination of two cytokines Huang Jian-feng, Huang Ji-feng, Zhang Wei-cai 2014, 18 (6):  829-834.  doi: 10.3969/j.issn.2095-4344.2014-06-002 Abstract ( 305 )   PDF (2003KB) ( 494 )   Save BACKGROUND:Bone marrow mesenchymal stem cells (BMSCs) can be induced to differentiate into neuron-like cells directionally. Accordingly, BMSCs can be used as seed cells theoretically in constructing tissue-engineered peripheral nerves. OBJECTIVE: Using combination of two cytokines to induce BMSCs differentiating into neuron-like cells directionally, and further to discusse its application in peripheral nerve injury. METHODS: BMSCs were isolated and purificated from the bone marrow of Wistar rats by using the differential adherence method. Basic fibroblast growth factor and epidermal growth factor were used to induce the BMSCs differentiating into neuron-like cells. The morphological change was observed and the neuronal specific markers were detected by immunohistochemistry technique. The morphological and immunohistological changes were also studied after the induce agent were removed. RESULTS AND CONCLUSION: With presence of morphological and immunohistochemical features of nerve cells induced by neurotrophic factors, BMSCs exhibited two or more processes that were interconnected as a meshwork; cell nucleus and nucleus could be observed with strong light refraction of cytoplasm. After immunohistochemical staining, neuroln specific enolase, neurofilament protein and synaptophysin protein positive cells were detected. A great amount of cells reversed to their original fibroblast-like morphology, and the expression of the three above-mentioned proteins decreased as the induce agent withdrawn. Our study showed that BMSCs can be induced to differentiate into neuron-like cells, but the transdifferentiation is a short-time reversible phenomenon. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effects of alpha-zearalanol on the osteogenic differentiation of mouse bone marrow mesenchymal stem cells Zou Bin, Zong Shao-hui, Zeng Gao-feng, Fang Ye, Gao Tai-hang 2014, 18 (6):  835-840.  doi: 10.3969/j.issn.2095-4344.2014.06.003 Abstract ( 346 )   PDF (832KB) ( 396 )   Save BACKGROUND: Bone marrow mesenchymal stem cells can differentiate into osteoblasts, and α-zearalanol can promote the osteogenic differentiation of mouse bone marrow mesenchymal stem cells. OBJECTIVE: To observe the effects of α-zearalanol on the osteogenic differentiation of mouse bone marrow mesenchymal stem cells. METHODS: Mouse bone marrow mesenchymal stem cells were isolated through whole bone marrow adherence method. Passage 3 cells were divided into non-induced group (cultured by the Iscove’s modified Dulbecco’s medium containing 10% fetal bovine serum and 1% penicillin and streptomycin), blank induced group (induced by the classic osteoblast-induced system with 0.1 μmol/L dexamethasone, 10 mmol/L β-glycerine sodium, and    50 μmol/L vitamin C), positive control induced group (induced by the classic osteoblast-induced system and   10-8 mol/L oestrogen), and low-, median-, high-concentration α-zearalanol groups (induced by the classic osteoblast-induced system and 10-7, 10-6, 10-5 mol/L α-zearalanol). The morphological changes of induced cells were observed by phase contrast microscopy, and the growth curves of cultured cells were plotted based on MTT results. The expression of alkaline phosphatase and osteocalcin were detected by ELISA. RESULTS AND CONCLUSION: In the non-induced group, cells showed long fusiform shape mostly, and contact inhibition was observed while the cells were close-packed. After induced, cells proliferated effectively, showed triangle, astero-form, polygon or irregular shape, and clumps and multilayer of cells became evident. Compared to both the blank induced group, the alkaline phosphatase activity and osteocalcin expression were significantly lower in the non-induced group, but higher in the positive control induced group and low concentration α-zearalanol group (P < 0.01). These findings indicate that α-zearalanol can promote the proliferation of bone marrow mesenchymal stem cells, and enhance the expression of alkaline phosphatase and osteocalcin expression, especially in the low concentration group. This demonstrates that low concentration α-zearalanol obviously enhances the osteogenic differentiation of mouse bone marrow mesenchymal stem cells in vitro. Up-regulating the release of alkaline phosphatase and osteocalcin may be one of the mechanisms by which α-zearalanol promotes osteogenic 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Autophagy-related gene Beclin-1 expression in neuron-like differentiation of human bone marrow mesenchymal stem cells Yang Yi, Ding Wen-jing, Dong Wan-li 2014, 18 (6):  841-846.  doi: 10.3969/j.issn.2095-4344.2014.06.004 Abstract ( 323 )   PDF (856KB) ( 579 )   Save BACKGROUND: Bone marrow mesenchymal stem cells have potential to self-renewal and multi-lineage differentiation. But after a long period of culture in vitro, the proliferation and differentiation capacities of bone marrow mesenchymal stem cells gradually loss, the mechanism underlying which is not clear now. OBJECTIVE: To observe the expression of autophagy-related gene Beclin-1 in differentiation from human bone marrow mesenchymal stem cells into neuron-like cells in vitro. METHODS: The changes of morphological characteristics of neuron-like cells differentiated from human bone marrow mesenchymal stem cells induced by epidermal growth factor were observed. The expression of neuron-specific enolase and glial fibrillary acidic protein in treated and untreated human bone marrow mesenchymal stem cells were detected using immunocytochemistry. The Beclin-1 protein expressions were detected by western blot before and after induction. RESULTS AND CONCLUSION: After being induced, human bone marrow mesenchymal stem cells presented classical neuron-like morphology; the expressions of neuron-specific enolase and glial fibrillary acidic protein were 78.7% and 8.1%, respectively. The expression of Beclin-1 protein was changed correspondingly during the induction, which increased after 30 minutes of induction and decreased gradually after 1 hour of induction. Human bone marrow mesenchymal stem cells could be induced into neuron-like cells in vitro by epidermal growth factor. Autophagy-related gene was highly expressed in the induction of early differentiation and the expression gradually reduced until it remained at a low level during the differentiation. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effect of different concentrations of uric acid on the neural differentiation of human bone marrow mesenchymal stem cells Song Qing-qing, Yu Ling-fan, Xu Li-li, Yang Nai-long 2014, 18 (6):  847-852.  doi: 10.3969/j.issn.2095-4344.2014.06.005 Abstract ( 394 )   PDF (484KB) ( 558 )   Save BACKGROUND: Uric acid as an endogenous antioxidant has garnered increasing attentions because of its anti-oxidation, anti-DNA damage and neuroprotective effects. OBJECTIVE: To observe the effect of uric acid at different concentrations on the neural differentiation of bone marrow mesenchymal stem cells. METHODS: Bone marrow mesenchymal stem cells were isolated, purified and cultured in vitro. The morphology change was observed. The third passages of bone marrow mesenchymal stem cells were induced to differentiate to neuron-like cells by induced liquid containing four different concentrations of uric acid (0 mmol/L as control group, 0.2 mmol/L, 0.4 mmol/L, 0.8 mmol/L) for 24 hours. Then, after second intervention for 1 hour, cells were detected by Nissl staining and specific markers were detected by immunohistochemistry method. RESULTS AND CONCLUSION: After induction, the cell body shrank, forming processes and connections. Nissl body was found in the cytoplasm. The positive rates of neuron-specific enolase were significantly higher in uric acid groups of different concentrations compared to the control group (P < 0.05); moreover, the positive rates of neuron-specific enolase were increased as the increase in concentrations of uric acid (P < 0.05). The positive rates of Nestin were decreased in uric acid groups of different concentrations compared to the control group(P < 0.05). After 4 hours of induction, cells fell off significantly. In a certain period of time, uric acid can promote differentiation of bone marrow mesenchymal stem cells into neuron-like cells in a certain concentration-dependent manner 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cells for epidermal and skin appendage regeneration Li Xian-song, Du Juan, Song Zhen-lan, Li Wei-ping 2014, 18 (6):  853-859.  doi: 10.3969/j.issn.2095-4344.2014.06.006 Abstract ( 355 )   PDF (2365KB) ( 419 )   Save BACKGROUND: Burned rat serum has been reported as an inducer that can induce differentiation of mesenchymal stem cells into epidermal cells. OBJECTIVE: To induce in vitro differentiation of bone marrow mesenchymal stem cells into epidermal cells that were transplanted alone or combined with inducers for the repair of skin wound defect and epidermal reconstruction. METHODS: Under aseptic environment, rat bone marrow was harvested to culture adherent cells using low-glucose Dulbecco’s modified Eagle’s medium. Culture cells at passage 4 were confirmed as mesenchymal stem cells by flow cytometry. Bone marrow mesenchymal stem cells were induced by Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 20% burned rat serum to differentiate into epidermal cells that were identified by immunohistochemistry. Wistar rat models of full-thickness skin wounds were prepared and divided into three groups. The 5-bromo-2-deoxyuridine-labeled autologous bone marrow mesenchymal stem cells and induced bone marrow mesenchymal stem cells were coated singly onto a rat model of burn wounds, and rat models of burn wounds with no treatment served as controls. Wound contraction and regeneration of epidermal cells and skin appendages were observed. RESULTS AND CONCLUSION: After isolation and culture of cells for 24 hours, a few of adherent cells grew as fibroblast-like cells with fusiform shape. At 16 days, cells completely covered the bottom of bottle, exhibiting a fish or reticular arrangement. After detection by flow cytometry, cells were cultured in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 20% burned rat serum, and fusiform-shaped cells gradually became round or oval cells. Flow cytometry analysis and immunocytochemistry results showed that cells expressed keratin, which were confirmed as epidermal cells. The results show that both the bone marrow mesenchymal stem cells transplantation alone or with necessary inductor is better for skin repair than natural healing, exhibiting a faster regeneration of skin and skin appendages. Bone marrow mesenchymal stem cells are deduced preliminarily to be involved in epidermis and hair follicle regeneration, thereby improving skin healing. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Morphological changes of goat bone marrow mesenchymal stem cells differentiating into fibrochondrocytes Su Xue-lian, Bao Guang-jie, Kang Hong, Liu Lin, Kong Nan-nan 2014, 18 (6):  860-865.  doi: 10.3969/j.issn.2095-4344.2014.06.007 Abstract ( 303 )   PDF (2456KB) ( 342 )   Save BACKGROUND: Our preliminary studies have shown that basic fibroblast growth factor can induce the differentiation of bone marrow mesenchymal stem cells into disc cells of the temporomandibular joint, and for basic fibroblast growth factor, 10 μg/L is superior to 5 μg/L in collagen synthesis. OBJECTIVE: To observe ultrastructural changes of bone marrow mesenchymal stem cells after being induced by different concentrations of basic fibroblast growth factor. METHODS: We cultured primary sheep bone marrow mesenchymal stem cells and selected passage 3 and 4 cells at good growth state. Bone marrow mesenchymal stem cells were stimulated with 5 and 10 μg/L basic fibroblast growth factor and their growth state was observed under inverted phase contrast microscope. Uninduced cells served as controls. The slides with cell crawling pieces were stained with Safranin O, picrosirius and type I collagen immunohistochemistry at days 7, 14 and 21, respectively. Simultaneously we collected the cells at day 21 to observe the ultrastructural changes of bone marrow mesenchymal stem cells. RESULTS AND CONCLUSION: After being induced with different concentrations of basic fibroblast growth factor, bone marrow mesenchymal stem cells were able to differentiate into disc cells of the temporomandibular joint; and after being induced with 10 μg/L basic fibroblast growth factor, cells were more like fibroblast-like cells of the temporomandibular joint disc. These findings indicate that bone marrow mesenchymal stem cells have morphological basis for differentiation to the fibroblast-like cells of the temporomandibular joint disc. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Extrapericardial adipose-derived stem cells differentiate into myocardial cells Zhang Jun, Zhang Yong, Lv Ji-yuan 2014, 18 (6):  866-871.  doi: 10.3969/j.issn.2095-4344.2014.06.008 Abstract ( 280 )   PDF (2008KB) ( 381 )   Save BACKGROUND: Adult stem cells are capable of proliferation and differentiation, which are an important part of tissue engineering. Although several studies have demonstrated that human adipose-derived stem cells obtained from elective liposuction procedures represent the characters of mesenchymal stem cells and can be induced to differentiate into myocardial cells, less is known about the characters of extrapericardial adipose-derived stem cells and their differentiation into myocardial cells. OBJECTIVE:To explore the culture of extrapericardial adipose-derived stem cells in vitro and their ability to differentiate into myocardial cells. METHODS:The extrapericardial adipose tissues were obtained and then isolated, cultured and passaged in vitro. The expressions of cell surface specific antigens (CD44 and CD90) were detected by immunofluorescence assay at different time periods, and the fluorescence intensity was compared. After induced and cultured with myocardium induction medium for an optimal period, α-actin and cardiac troponin T expressions were detected by immunocytochemical detection and the phenotypic identifications were performed. RESULTS AND CONCLUSION: During the in vitro culture, extrapericardial adipose-derived stem cells adhered to culture flask wall and proliferated strongly. The cells appeared to be shuttle-shaped and exhibit a vortex alignment. They were positive for CD44 and CD90, and the fluorescence intensity reached the peak at passages 3 and 4, but negative for CD34 and CD54. After induced and cultured with myocardium induction medium, the differentiated cells were muscle-like cells, the expression of α-actin and cardiac troponin T were positive, and the number of cells reached the peak at passages 3 and 4. These findings suggested that the extrapericardial adipose-derived stem cells from human represent the morphological and cellular immunological characters of mesenchymal stem cells, and they can be induced to differentiate into myocardial cells. In addition, there is the best induction period. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Three-dimensional spherical culture of adipose-derived stem cells Wang Chan, Dai Ying, Guo Yong-long, Yang Yan, Liu Qing, Chen Jian-su 2014, 18 (6):  872-879.  doi: 10.3969/j.issn.2095-4344.2014.06.009 Abstract ( 509 )   PDF (2692KB) ( 703 )   Save BACKGROUND:Many types of mammalian cells aggregate and display three-dimensional multicellular spheroids when they are in normal physiological conditions. In order to observe and explore cellular natural states, many researchers try to use spherical cell culture in vitro, a common three-dimensional culture pattern. OBJECTIVE:To use three different methods for spherical culture in vitro of adipose-derived stem cells and to observe their biological features. METHODS: Adipose-derived stem cells were confirmed by the analysis of the markers for cell phenotypes as well as adipogenic and osteogenic differentiation potential assays. Three different methods of sphere cultures were used as follows: (1) ultra low attachment culture; (2) hanging-drop culture and (3) Eppendorf tube culture. The sphere formation was compared among above three methods. We used Imagej to calculate mean areas of these spheres. And we used Viability/Cytotoxicity Assay Kit for Animal Live & Dead Cells to detect their vitality. RESULTS AND CONCLUSION: (1) Adipose-derived stem cells were confirmed by the analysis of the markers for cell phenotypes, CD29, CD44, CD59 were positive, as well as adipogenic and osteogenic differentiation potential assays were positive. The conventional monolayer cultures of adipose-derived stem cells showed spindle and cloning growth within three passages. (2) Ultra low attachment culture, hanging-drop culture, Eppendorf tube culture all could elicit adipose-derived stem cells spherical growth. However, spherical size, shape and uniformity differed depending on cell numbers, culture time and spherical culture methods. The ultra low attachment culture was comparatively difficult to control spherical shape and uniformity of adipose-derived stem cells. But hanging-drop culture and Eppendorf tube culture were able to form even cell spheres. (3) Spherical formation of adipose-derived stem cells using our three methods displayed good cell vitality. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Proliferation of hematopoietic stem cells differentiated from embryonic stem cells via sustained Wnt pathway activation Lin Fang, Jin Jing-jun, Zhang Tao, Ji Bin-feng, Chen Yong 2014, 18 (6):  880-887.  doi: 10.3969/j.issn.2095-4344.2014.06.010 Abstract ( 499 )   PDF (2429KB) ( 711 )   Save BACKGROUND:A variety of embryonic stem cells induction and differentiation systems have been established so far, while the research that promotes embryonic stem cells to differentiate into hematopoietic stem cells is still at an initial stage, and the induction efficiency needs to be improved. OBJECTIVE: To active the Wnt/β-catenin signal pathway in mouse embryonic stem cells with exogenous win3a as an inducer, and then to observe whether the activation of this pathway will promote the directional differentiation of embryonic stem cells into hematopoietic progenitor cells. METHODS:The ES-E14TG2a mouse stem cells were cultured with the exogenous wnt3a (100 µg/L) for 21 days, the content of β-catenin was tested by cell immunofluorescence and western blot, and expression of Wnt downstream target gene was detected by quantitative reverse transcription PCR to determine the activation of Wnt/β-catenin signal pathway. Single-layer adherent culture method was used to induce the directional differentiate of above-mentioned cells into hematopoietic stem cells, and detection of hematopoietic development associated surface marker CD34+/Sca-1+ was achieved by flow cytometry; meanwhile, the expression of hematopoietic associated gene was measured by quantitative reverse transcription PCR. RESULTS AND CONCLUSION: We found that β-catenin accumulated in ES-E14TG2a mouse stem cells after cultured with wnt3a (100 µg/L) for 21 days; the expressions of Wnt downstream target genes such as Pitx2, Frizzled, Sox17 and Oct4 showed the different degrees in increase, meaning the activation of Wnt/β-catenin signal pathway. Furthermore, during the time that we used single-layer adherent culture method to induce hematopoietic stem cells, the CD34+/Sca-1+ cells accounted for 20.2% of total cells at day 14, and control cells only accounted for 11.9%. Again, expression quantity of hematopoietic associated gene BMP4, FLK2 and CD34 increased while Smad5 was suppressed significantly. Our data suggest that sustaining action by wnt3a will active Wnt/β-catenin signal pathway, and also promote the directional differentiation of ES-E14TG2a mouse stem cells into hematopoietic progenitor 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Stem cell tumorigenicity in Balb/c nude mice Nie De-zhi, Wan Ying, Ben Liang, Wang Ying-jun, Liu Xiang-zhu, Wang Li-hui, Li Chao, Zhang Shi-dong 2014, 18 (6):  888-893.  doi: 10.3969/j.issn.2095-4344.2014.06.011 Abstract ( 1137 )   PDF (1751KB) ( 1293 )   Save BACKGROUND:Stem cell tumorigenicity is a practical issue concerning stability in the clinical application of stem cells. Therefore, it is particularly important to clear whether stem cells have tumorigenic ability or not. Nude mice occupy an increasingly important position in oncology, immunology, and safety evaluation of drugs and biological products. OBJECTIVE: To observe the tumorigenicity of neural stem cells and mesenchymal stem cells in Balb/c nude immunodeficient mice. METHODS: Balb/c nude mice were randomly divided into control group, negative group, positive group, neural stem cell group and mesenchymal stem cell group. HepG-2 cells, RPE cells, passage 4 neural stem cells and mesenchymal stem cells were injected subcutaneously into nude mice from different groups, respectively. After 12 weeks of injection, anatomical observation was performed to detect the tumor formation in the transplantation site. Meanwhile, soft agar colony formation assay was applied to investigate neural stem cell and mesenchymal stem cell clone in vitro. RESULTS AND CONCLUSION:After 12 weeks of injection, the tumorigenicity study results showed that no tumor developed in the transplantation site in the control group, negative group, neural stem cell group and mesenchymal stem cell group. Histopathologic examinations also showed no abnormality in these groups. Soft agar colony formation assay showed in soft agar resistance medium, neural stem cells and mesenchymal stem cells did not exhibit clone ability. These findings indicate that neural stem cells and mesenchymal stem cells undergoing short-term passages have no tumorigenic growth. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Correlation between Bmi-1 and clinicopathological features of colorectal cancer Huang Zhe 2014, 18 (6):  894-899.  doi: 10.3969/j.issn.2095-4344.2014.06.012 Abstract ( 356 )   PDF (597KB) ( 885 )   Save BACKGROUND: Bmi-1 (B-cell-specific moloney murineleukemia virus insertion site), a colorectal cancer stem cell gene is of great significance in the regulation of telomere reverse transcriptase activity and transcriptional status as well as cell aging and carcinogenesis. OBJECTIVE: To investigate the correlation between Bmi-1 expression and clinicopathological changes of colorectal cancer. METHODS: The expression of Bmi-1 mRNA in carcinoma cells was detected in 50 cases of colorectal cancer using quantitative real-time PCR. The expression of Bmi-1 protein in cancer tissue was detected by hematoxylin-eosin staining and immunohistochemical staining SP method. RESULTS AND CONCLUSION: Sex, age, and tumor size showed no different effects on the expression of Bmi-1 mRNA. The expression of Bmi-1 protein showed no significant difference in patients who were different in sex, age, tumor size and Duke staging of tumor. The expressions of Bmi-1 mRNA and protein in patients with lymph node metastasis and depth of invasion deeper than the serosa were much higher than those in patients with no lymph node metastasis and depth of invasion shallower than the serosa (P < 0.05). The expression of Bmi-1 mRNA was much higher in patients with higher Duke staging cancer (P < 0.05). The expression of Bmi-1 mRNA and protein are closely related to different pathological features of colorectal cancer, such as metastasis, infiltration, and Duke staging. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effect of adipose-derived mesenchymal stem cell transplantation on inflammatory response and ventricular remodeling after myocardial infarction Fan Yan, Wang Jian-jun, Wei Feng, Fan Xiao-hai, Ma Ai-qun 2014, 18 (6):  900-905.  doi: 10.3969/j.issn.2095-4344.2014.06.013 Abstract ( 370 )   PDF (836KB) ( 778 )   Save BACKGROUND: Whether adipose-derived mesenchymal stem cells are able to exert immunomodulatory effects in the treatment of myocardial infarction, as well as the best time, is less reported. OBJECTIVE: To observe the effect of adipose-derived mesenchymal stem cells on inflammatory reaction and ventricular remodeling after myocardial infarction, and to explore the possible mechanisms of adipose-derived mesenchymal stem cells for the treatment of myocardial infarction. METHODS: Enzyme digestion method was employed to isolate and culture rat adipose-derived mesenchymal stem cells. By ligation of the left anterior descending coronary artery, we established animal models of myocardial infarction in 40 rats. The rats were randomly divided into four groups: sham group, control group (injected high-glucose Dulbecco’s modified Eagle’s medium), 3-hour transplantation group (transplanted adipose-derived mesenchymal stem cells after 3 hours of myocardial infarction), 7-day transplantation group (transplanted adipose-derived mesenchymal stem cells after 7 days of myocardial infarction). After 14 days of operation, the levels of tumor necrosis factor-α and interleukin-10 in the plasma were detected by enzyme linked immunosorbent assay. After 28 days of operation, the left ventricular end diastolic diameter, left ventricular end systolic diameter, left ventricular ejection fraction and left ventricular fractional shortening were measured by echocardiography. RESULTS AND CONCLUSION: Compared with the control group, in the 3-hour transplantation group and 7-day transplantation group, the levels of tumor necrosis factor-α were significantly lower (P < 0.01), and the levels of interleukin-10 were significantly higher (P < 0.01) at postoperative 14 days; the left ventricular end diastolic diameter and left ventricular end systolic diameter in the two transplantation groups were also significantly smaller (P < 0.05), but left ventricular ejection fraction and left ventricular fractional shortening were significantly elevated (P < 0.05), which was more apparent in the 3-hour transplantation group than the 7-day transplantation group. Adipose-derived mesenchymal stem cells transplantation in acute phase of myocardial infarction can suppress the inflammatory response, regulate the cytokine network equilibrium, and thus delay ventricular remodeling and improve cardiac function. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cell transplantation suppresses emphysema-induced inflammation and apoptosis Zhao Xiao-jian, Lu Cai-ping, Chu Wei-wei, Zhen Qiang, Tan Guo-liang, Zhang Ya-xiao, Wang Ren-feng, Zhang Bing, Liu Jia-bao 2014, 18 (6):  906-911.  doi: 10.3969/j.issn.2095-4344.2014.06.014 Abstract ( 350 )   PDF (850KB) ( 452 )   Save BACKGROUND: Bone marrow mesenchymal stem cells transplantation can change the surrounding microenvironment through paracrine mechanisms, and can be employed for treatment of serious damage to lung function through the promotion of angiogenesis, inhibition of apoptosis and maintaining functional stability of autonomic nervous system. OBJECTIVE: To observe the inflammatory reaction in experimental emphysema and inhibition of apoptosis through bone marrow mesenchymal stem cells transplantation. METHODS: Twenty-four Wistar female rats were randomly divided into three groups: healthy control group, model group and experimental group. In the latter two groups, smoking and endotracheal instillation of porcine pancreatic elastase were performed to establish emphysema models. After modeling, bone marrow mesenchymal stem cells were injected via tail vein in the experimental group. Pathological changes of the lung, the level of tumor necrosis factor-alpha and cell number in the bronchoalveolar lavage fluid as well as apoptotic index in lveolar walls were detected after cell transplantation. RESULTS AND CONCLUSION: In the model and experimental groups, pathological changes of lung tissues were observed to different extent. The lung pathological changes were slighter in the experimental group than the model group (P < 0.01). The level of tumor necrosis factor-alpha and apoptotic index in lung tissue were lower in the experimental group than the model group (P < 0.01). These findings indicate that bone marrow mesenchymal stem cells can improve emphysema pathologically through inhibition of inflammatory response and apoptosis in experimental emphysema. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Mononuclear cells isolated from mobilized bone marrow differentiate into vascular endothelial cells and cardiomyocyte-like cells Yao Wei, Zhang Xiu-lan, Yao Ying, Wang Wei-shu 2014, 18 (6):  912-918.  doi: 10.3969/j.issn.2095-4344.2014.06.015 Abstract ( 362 )   PDF (2279KB) ( 454 )   Save BACKGROUND: It is controversial whether bone marrow mobilization can retain in cardiac injured position in congestive cardiomyopathy or differentiate into cardiomyocytes and vascular endothelial cells. OBJECTIVE: To study the effects of granulocyte colony stimulating factor (G-GSF) on myocardium and angiogenesis in rats with congestive cardiomyopathy. METHODS: Fifty Wistar rats with heart failure caused by adriamycin-induced cardiomyopathy were divided into heart failure group (n=20) treated with normal saline and bone marrow mobilization (n=30) treated with subcutaneous injection of recombinant human G-GSF. Ten rats from the bone marrow mobilization were killed at day 6 of mobilization, and myocardial tissue was taken for CD43 immunofluorescent staining. Blood samples were taken from the rat tail in each group before and 5 days after treatment to count total number of white blood cells and percentage of mononuclear cells. Meanwhile, mononuclear cells extracted from the peripheral blood were used for flow cytometry detection. At day 5 after treatment, bromodeoxyuridine (BrdU, 50 mg/kg) was successively given to all rats for 4 weeks before they were sacrificed. Myocardial tissues were taken to determine the homing of mononuclear cells and evaluate differentiation of mononuclear cells into cardiomyocytes and vascular endothelial cells using BrdU staining, BrdU/myosin heavy chain double staining, and BrdU/actin double staining. Hematoxylin-eosin staining was used for determination of blood vessel density. RESULTS AND CONCLUSION: G-CSF mobilization increased the number of mononuclear cells that was significantly higher than before treatment (P < 0.05). Flow cytometry showed that the number of CD34-positive mononuclear cells in the peripheral blood was higher in the bone marrow mobilization than in the heart failure group (P < 0.05). Myocardial CD34 immunofluorescence showed that the heart failure group was negative and the bone marrow mobilization group was positive. In the bone marrow mobilization group, the myocardial tissue was positive for BrdU staining, BrdU/myosin heavy chain double staining and BrdU/actin double staining, while vascular endothelial cells in the region of myocardial injury was positive for BrdU; conversely, the heart failure group was negative. The density of blood vessels in the bone marrow mobilization group was significantly higher than that in the heart failure group (P < 0.001). These findings indicate that bone marrow mobilization increases the number of mononuclear cells, and these cells are homing to myocardial injury, thereby playing a repair role in the myocardium and vascular tissue of heart failure rats with congestive cardiomyopathy. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effect of bone marrow mesenchymal stem cells transplantation on the expression of CD163 and interleukin-10 in rats with acute hepatic liver failure Yuan Shu-fang, Hu Lan-ying, Jiang Tao, Sun Li-hua, Zheng Rong-jiong, Zhao Jin-yan, Zhang Yue-xin 2014, 18 (6):  919-925.  doi: 10.3969/j.issn.2095-4344.2014.06.016 Abstract ( 273 )   PDF (2255KB) ( 536 )   Save BACKGROUND: Studies have shown that bone marrow mesenchymal stem cells have the ability to persistently generate hepatocytes and biliary cells, and thus in the repair process of liver injury, replenish the reduced number of hepatocytes due to damage and participate in damaged liver structure. OBJECTIVE: To investigate the effect of bone marrow mesenchymal stem cells transplantation on acute liver failure and the expression of CD163 and interleukin-10 in rat serum and liver tissue. METHODS: D-galactose and lipopolysaccharidewere used to make acute liver failure models in 60 Sprague-Dawley rats. Then, the rats were divided into control group and transplantation group. Bone marrow mesenchymal stem cells at passage 3 were injected through tail vein in the transplantation group, and normal saline was injected in the control group. After transplantation 24, 120, 168 hours, serum samples and liver tissues were collected. RESULTS AND CONCLUSION: After transplantation 120 and 168 hours, the serum alanine transaminase and aspartate aminotransferase activities of the transplantation were significantly lower than those of the control group (P < 0.05). In the transplantation group the apoptotic index was still lower compared with the control group, and the difference was significant (P < 0.05). The levels of CD163 and interleukin-10 in the serum and liver tissue in the transplantation group were decreased significantly compared with the control group (P < 0.05). The results suggested that there were highly significant correlations between CD163 and interleukin-10 (P < 0.01). Bone marrow mesenchymal stem cell transplantation has a therapeutic effect on acute liver failure rats. CDl63 and interleukin-10 play a very important role in the pathogenesis of acute liver failure, which can be used as sensitive serum marker proteins for diagnosis and prognosis of acute liver failure. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Different sources of mesenchymal stem cells in the treatment of liver fibrosis in rats Qu Xin, Wang Xin-chao, Han Lu, Zhang Hai-chao 2014, 18 (6):  926-931.  doi: 10.3969/j.issn.2095-4344.2014.06.017 Abstract ( 347 )   PDF (1956KB) ( 452 )   Save BACKGROUND: In recent years, studies have shown that mesenchymal stem cells can secrete various growth factors, and has certain application prospect in the treatment of liver fibrosis. OBJECTIVE: To explore the different sources of mesenchymal stem cells in the treatment of liver fibrosis in rats. METHODS: Fifty Sprague-Dawley rats were given intraperitoneal injection of carbon tetrachloride to construct the model of liver fibrosis. Another 10 normal Sprague-Dawley rats were used as normal controls. Model rats were randomly divided into five groups, 10 rats in each group, including model control group, normal saline group, bone marrow mesenchymal stem cell group, umbilical cord mesenchymal stem cell group, and umbilical cord blood mesenchymal stem cell group. After 8 weeks of modeling, different sources of mesenchymal stem cells at a density of 2×106 were injected via tail vein into model rats. After 12 weeks, the rats were killed, and serum alanine aminotransferase, aspartate aminotransferase and albumin levels were detected as well as the expression of type I collagen and glial fibrillary acidic protein in liver tissue was detected by fluorescence quantitative PCR method. RESULTS AND CONCLUSION: We have successfully established the SD rat model of hepatic fibrosis. Bone marrow mesenchymal stem cells aggravated hepatic fibrosis, umbilical cord blood and umbilical cord mesenchymal stem cells could reduce the level of hepatic fibrosis in rats. Umbilical cord mesenchymal stem cells had the most obvious effect that significantly reduced expressions of alanine aminotransferase, aspartate aminotransferase and type I collagen and glial fibrillary acidic protein. The results showed different sources of mesenchymal stem cells have different effects on rat fibrosis. Bone marrow mesenchymal stem cells aggravate hepatic fibrosis, and umbilical cord and umbilical cord blood stem cells can alleviate hepatic fibrosis 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Therapeutic applications of umbilical cord mesenchymal stem cells in Parkinson’s disease Wang Yan, Zhao Xin-li, Zhang Jun-yan, Tan Jun 2014, 18 (6):  932-937.  doi: 10.3969/j.issn.2095-4344.2014.06.018 Abstract ( 643 )   PDF (800KB) ( 1005 )   Save BACKGROUND: Animal experiments have repeatedly verified that umbilical cord stem cell transplantation can improve the rotational behavior of rats with Parkinson’s disease, with lower immune rejection response. OBJECTIVE: To evaluate the therapeutic effects of human umbilical cord mesenchymal stem cell transplantation in the treatment of Parkinson’s disease using the Unified Parkinson’s Disease Rating Scale. METHODS: Fifteen patients with Parkinson’s disease were enrolled, including 8 males and 7 females, aged 52-76 years. Hoehn & Yahr staging was 3-5. After informed consent was obtained from puerperae and the procedure was approved by the hospital ethics committee, full-term maternal umbilical cord was aseptically collected, to culture umbilical cord mesenchymal stem cells. All patients were hospitalized, and treated with human umbilical cord mesenchymal stem cell transplantation via carotid artery puncture. Neurological function of patients was assessed using a comprehensive rating scale for Parkinson’s disease before and 1 month after cells transplantation. Higher score indicated more severe neurological deficits. RESULTS AND CONCLUSION: Fifteen patients entered the result analysis. Compared with before transplantation, 15 patients showed significantly lowered scores on the Unified Parkinson’s Disease Rating Scale at 1 month after transplantation (P < 0.05). The improvement was mainly concentrated in the tremor and rigidity, but bradykinesia and unstable position were not improved. Graft versus host disease did not occur in all 15 cases. These findings indicate that umbilical cord blood mesenchymal stem cell transplantation for treating Parkinson’s disease has obvious curative effects and significantly improves neurological functions. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effects of human bone marrow cells-derived extracellular matrix on the proliferation of human periodontal ligament stem cells Wang A-xian, Tang Li, Liang Yuan, Ji Hai-ning, Wu Jun-jie, Ding Yin 2014, 18 (6):  938-943.  doi: 10.3969/j.issn.2095-4344.2014.06.019 Abstract ( 364 )   PDF (993KB) ( 414 )   Save BACKGROUND: Human bone marrow cells-derived extracellular matrix can promote proliferation of human periodontal ligament stem cells and maintain stem cell properties. OBJECTIVE: To preliminarily investigate the effect of human bone marrow cells-derived extracellular matrix on the proliferation of human periodontal ligament stem cells. METHODS: Human periodontal ligament stem cells and bone marrow cells were separately derived from human periodontal tissue and jaw bone marrow, and human bone marrow cells-derived extracellular matrix was prepared. Human periodontal ligament stem cells were cultured and purified using limited dilution cloning method, and transmission electron microscope was used for ultrastructure observation. Human periodontal ligament stem cells at passage 2 were cultured with human bone marrow cells-derived extracellular matrix and normal culture medium (control group). The cell counting kit-8 and flow cytometry were used to determine the proliferation potential of human periodontal ligament stem cells cultured on human bone marrow cells-derived extracellular matrix. RESULTS AND CONCLUSION: Compared with the control group, human periodontal ligament stem cells cultured on human bone marrow cells-derived extracellular matrix had a superior capacity of proliferation (P < 0.05), and the cells met their morphological and biological characteristics, and grew in good conditions. Human bone marrow cells-derived extracellular matrix is a promising matrix for large-scale expanding human periodontal ligament stem cells for future use in stem cell-based therapy. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Isolation and culture of human amniotic mesenchymal stem cells: proper digestion time and concentrations of trypsin and collagenase Zhang Hui-juan, Cong Shan, Liang Mei-ping, Liu Jun-ping, Huang Li-gang, Song Jin, Cao Gui-fang 2014, 18 (6):  944-949.  doi: 10.3969/j.issn.2095-4344.2014.06.020 Abstract ( 455 )   PDF (2297KB) ( 363 )   Save BACKGROUND: Extraction methods of human amniotic mesenchymal stem cells are inconsistent in the number of cells. OBJECTIVE: To explore the optimal method to in vitro isolate and culture human amniotic mesenchymal stem cells. METHODS: Under sterile conditions, full-term cesarean fetal amniotic membrane was cut into pieces, then to isolate human amniotic mesenchymal stem cells by seven methods in four experiments. In experiment 1, human amniotic mesenchymal stem cells were isolated by the following three methods: (1) 0.05 g/L trypsin digestion for 10 minutes followed by 0.75 g/L collagenase digestion for 60 minutes; (2) 0.75 g/L collagenase I for 120 minutes; (3) co-digestion with 0.05 g/L trypsin and 0.75 g/L collagenase for 60 minutes. In experiment 2, the samples were digested with 0.05 g/L trypsin digestion for 30 minutes followed by 0.75 g/L collagenase digestion for 30 minutes. In experiment 3, the samples were digested by two methods: (1) 0.05 g/L trypsin digestion for 30 minutes×2, followed by 0.75 g/L collagenase digestion for 60 minutes; (2) 0.05 g/L trypsin digestion for 40 minutes×2, followed by 0.75 g/L collagenase digestion for 60 minutes. In experiment 4, the samples were digested with 0.05 g/L trypsin digestion for 30 minutes×2, followed by 1 g/L collagenase digestion for 60 minutes. Following morphology observation under a microscope, we studied the most suitable method for isolating human amniotic mesenchymal stem cells. RESULTS AND CONCLUSION: Digestion with 0.05 g/L trypsin for 30 minutes twice followed by 1 g/L of collagenase digestion of 60 minutes was the most suitable isolation and culture condition in vitro. Cells became elongated fusiform or star-shaped with rich cytoplasm, and nuclei were round with 1-3 nuts. We can harvest the most number of human amniotic mesenchymal stem cells using the method described in experiment 4. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Stromal cell derived factor 1 effects on migration of endogenous neural stem cells Su Wen, Ding Peng, Wang Jin-kun, Zhang Hao, Mu Lin-jie, Wang Bo, Liu Jing-chuan, Gong Guang-hui, Wang Chong-qian 2014, 18 (6):  950-955.  doi: 10.3969/j.issn.2095-4344.2014.06.021 Abstract ( 299 )   PDF (2521KB) ( 372 )   Save BACKGROUND:Stromal cell derived factor 1 in chemotactic migration of endogenous neural stem cells plays a very important role, but the specific migration mechanism is unclear OBJECTIVE: To observe the effects of exogenous stromal cell derived factor 1 on chemotactic migration and proliferation of neural stem cells in the rat hippocampus METHODS:Exogenous stromal cell derived factor 1 (5 μL, 500 μ/L) was injected into the hippocampus of Sprague-Dawley rats to establish animal models. Brain tissues were taken after days 3, 7, 14 and 21 of perfusion to prepare paraffin sections. Thereafter, nestin expression in the injection region and hippocampus was detected using immunohistochemical method. Experimental control and blank control groups were set. RESULTS AND CONCLUSION: Paraffin section immunohistochemical results displayed the number of nestin-positive cells in the injection and the hippocampus was gradually increased. At 3 and 7 days, nestin expression was a little and increased at 14 days, forming a migration tendency to the injection region. At 21 days, there were more nestin-positive cells in the injection area and hippocampus. However, there were no changes as above in the experimental control and blank control groups. The results showed that exogenous stromal cell derived factor 1 may induce the proliferation of neural stem cells in the hippocampus and may be involved in chemotactic migration of endogenous 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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The role of Toll-like receptor 1 in the immunoregulatory regulation of Th17 cells by mesenchymal stem cells Guo Zhen-xing, Chen Zhen-ping 2014, 18 (6):  956-961.  doi: 10.3969/j.issn.2095-4344.2014.06.022 Abstract ( 284 )   PDF (1815KB) ( 338 )   Save BACKGROUND: Our previous studies have shown that mesenchymal stem cells play an immunomodulatory role in Th17 cells, but the mechanism of action remains to be elucidated, therefore, to explore the role of Toll-like receptor 1 in which will provide possible experimental basis for the future potential of cell therapy strategies. OBJECTIVE: To investigate the role of Toll-like receptor 1 in the immunoregulatory function of mesenchymal stem cells on Th17 cells. METHODS: Separation of adherent bone marrow-derived human embryonic sources of mesenchymal stem cells, immunomagnetic separation of normal CD4+ T cells. CD4+ T cells were cultured alone or in combination with mesenchymal stem cells co-cultured 4d. Real-time quantitative polymerase chain reaction assay interleukin-17, Toll-like receptor 1 expression levels related genes; number of Th17 cells by flow cytometry. Bone marrow mesenchymal stem cells from human embryos were separated using adherence method, and co-cultured with human CD4+ T cells from healthy donors using immunomagnetic separation method for 4 days. The expression of interleukin-17 and Toll-like receptor 1 mRNA was detected by real-time PCR, and the number of Th17 cells was observed by flow cytometry. RESULTS AND CONCLUSION: Toll-like receptor 1 mRNA was expressed in both CD4+ T cells and mesenchymal stem cells. Level of interleukin-17 mRNA was significantly higher in mesenchymal stem cells co-cultured with CD4+ T cells than in CD4+ T cells cultured alone (relative value 3.59±0.11 vs. 1.14±0.08, P < 0.01). Consistent with the expression of interleukin-17 mRNA, increased level of Toll-like receptor 1 mRNA was detected in mesenchymal stem cells co-cultured with CD4+ T cells compared with CD4+ T cells cultured alone (relative value 6.07±1.79 vs. 1.53±0.63, P < 0.01). Furthermore, Flow cytometry analysis showed that the percentage of Th17 cells in the mesenchymal stem cells co-cultured with CD4+ T cells was significantly higher than that in CD4+ T cells cultured alone, (4.53±1.27)% vs. (2.39±0.80)% (P < 0.01). Toll-like receptor 1 might be involved in the immunoregulatory regulation of Th17 cells by 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Impact of perioperative cardiovascular risk factors on bone marrow progenitor cells Zhang Lin, Gao Chang-qing, Wang Rong, Li Li-bing, Cheng Nan, Yao Ming-hui 2014, 18 (6):  962-967.  doi: 10.3969/j.issn.2095-4344.2014.06.023 Abstract ( 287 )   PDF (801KB) ( 361 )   Save BACKGROUND: Cell therapy by the implantation of autologous bone marrow cells has been used for the treatment of ischemic heart diseases in clinical trials for decade. However, as the outcomes of cell transplantation obviously vary among patients, it is essential to identify the risk factors that may influence the level and function of progenitor cells in bone marrow, in order to identify the patients who would benefit the most from this treatment. OBJECTIVE: To observe the impact of perioperative cardiovascular risk factors on number and function of bone marrow progenitor cells from patients undergoing coronary artery bypass grafting surgery. METHODS: We collected clinical and laboratory data from 44 patients scheduled to undergo sternotomy for coronary artery bypass grafting procedures. Bone marrow was aspirated from the sternum during the operation and bone marrow mononuclear cells were isolated by density centrifugation with Ficoll lymphoprep and then detected using trypan blue exclusion method. Levels of progenitor cells in bone marrow were evaluated using flow cytometry. Function of bone marrow progenitor cells were assessed by clonogenic and migration assays. RESULTS AND CONCLUSION: We assessed the number of bone marrow mononuclear cells out of 20 mL bone marrow in duplicate samples from patients with coronary heart disease scheduled for coronary artery bypass grafting that was (10-89)×106 cells with over 95% activity. A negative correlation was observed between the number of bone marrow mononuclear cells and the age (n=44, r=-0.788, P=0.001). Levels of CD34+, CD133+, and CD34+CD133+ cells in bone marrow mononuclear cells was (0.94±0.39)%, (0.46±0.28)%, and (0.53±0.26)%. Levels of CD34+ cells and CD133+ cells in patients with diabetes were significantly lower than those in patients without diabetes. Female, advanced age and poor heart function were related with reduced colony-forming ability of progenitor cells. A positive correlation was observed between level of CD34+ cells and migration ability of bone marrow mononuclear cells. The results show that by density gradient centrifugation, we can harvest a sufficient number of bone marrow mononuclear cells in the treatment for ischemic heart disease. Age, gender, diabetes, heart function are correlated with bone marrow mononuclear cell number and functions. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cells for osteoarthritis: its possibility and future Zhang Ping-ping, Xiang Chuan 2014, 18 (6):  968-973.  doi: 10.3969/j.issn.2095-4344.2014.06.024 Abstract ( 682 )   PDF (613KB) ( 964 )   Save BACKGROUND: Nowadays treatment of osteoarthritis with drugs is not ideal. In recent years, more and more scientists try to use stem cells to treat osteoarthritis. OBJECTIVE: To explore the stem cell treatment for osteoarthritis in order to promote its clinical application and find out its difficulties. METHODS: A computer-based online search of PubMed database and CNKI database between March 1998 and October 2013 was performed to search related articles with the key words of “stem cell, osteoarthritis, bone metabolism, bone marrow mesenchymal stem cells” in English or in Chinese, respectively. The word “AND” was used for the connection between the word retrieval. Literatures related to stem cells treatment for osteoarthritis were selected; in the same field, the articles published lately in authoritative journals were preferred. RESULTS AND CONCLUSION: A total of 79 literatures were primarily selected, and 50 documents were involved for summary according to inclusion criteria. The stem cell treatment for osteoarthritis is realized mainly by promoting the repair of cartilage tissue. Commonly used methods are stem cell transplantation and induced differentiation of stem cells. Stem cell treatment has broad application prospects for the treatment of osteoarthritis. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Neural stem cell transplantation for central nervous system diseases via the cerebrospinal fluid Zhang Bao-hua, Qiu Fu-cheng, Dong Ci1, Han Rui, Zhang Yong-zhi, Liu Hui-miao,Xie Bing-chuan, Zhang Li-na, Wang Wen-ting, Wang Yan-yong, Zhang Zhen-qing, Gu Ping,Yan Bao-yong 2014, 18 (6):  974-978.  doi: 10.3969/j.issn.2095-4344.2014.06.025 Abstract ( 320 )   PDF (666KB) ( 460 )   Save BACKGROUND: Currently, neural stem cell transplantation can be performed through three main approaches: local lesions, blood circulation, and cerebrospinal fluid. OBJECTIVE: To review the transplantation of neural stem cells or neural precursor cells via the cerebrospinal fluid in the treatment of central nervous system diseases. METHODS: A computer-based search of PubMed and CHKD databases was performed to retrieve articles concerning transplantation of neural stem cells via the cerebrospinal fluid, and its application and therapeutic mechanism in the treatment of central nervous system diseases in both animal experiment and clinic study published from 2000 to 2009. RESULTS AND CONCLUSION: It is suitable for neural stem cell survival, proliferation, and differentiation in the cerebrospinal fluid. Transplantation of neural stem cells via the cerebrospinal fluid is effective and feasible to treat central nervous system diseases. However, some problems have not been solved, such as the source of neural stem cells, the optimal time window and cell dose, the safety and the long-term effect. Further studies are needed to pave the way for the intrathecal injection of neural stem cells in the treatment of central nervous system diseases. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effects of epimedium on osteogenic differentiation of bone marrow mesenchymal stem cells Li Hui-zhen, Li Meng, Li Rui-yu, Wu Li-ping 2014, 18 (6):  979-984.  doi: 10.3969/j.issn.2095-4344.2014.06.026 Abstract ( 386 )   PDF (851KB) ( 509 )   Save BACKGROUND: Epimedium has been used in the treatment of osteoporosis and repair of bone defects. OBJECTIVE: To explore the effects of epimedium on the osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS: A database search was performed to retrieve literatures addressing epimedium effects on the osteogenic differentiation of bone marrow mesenchymal stem cells. Then, the papers meeting the criteria were selected for in-depth analysis. During the osteogenic differentiation induced by epimedium, alkaline phosphatase, osteocalcin, osteopontin, transforming growth factor β, bone morphogenetic proteins, osteocalcin, bone sialoprotein were detected in the bone marrow mesenchymal stem cells to understand the underlying mechanism of epimedium in proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells. RESULTS AND CONCLUSION: Epimedium effects on bone marrow mesenchymal stem cells are shown in a dose-dependent manner. During the early induction, icariin can increase cell phosphatase activity; in the late induction, icariin can increase calcified nodules, promote osteocalcin secretion, significantly improve the expressions of transforming growth factor β1, bone morphogenetic protein 2, insulin-like growth factor-1, osteopontin and bone sialoprotein. Epimedium, which can be used as an excellent osteoinductive factor, improves the proliferation and osteogenic 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 全文链接： Figures and Tables | References | Related Articles | Metrics