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01 January 2015, Volume 19 Issue 1 Previous Issue    Next Issue

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Hydrogen peroxide influences embryonic stem cell transplantation in the treatment of myocardial infarction Fan Zhi-gang, Kong Chun-yan 2015, 19 (1):  1-6.  doi: 10.3969/j.issn.2095-4344.2015.01.001 Abstract ( 300 )   PDF (913KB) ( 753 )   Save BACKGROUND: Specific mechanism of stem cell transplantation for treatment of myocardial infarction has been not very clear. OBJECTIVE: To investigate the effect of H2O2-pretreated mouse embryonic stem cells in vitro for treatment of myocardial infarction. METHODS: Twenty-four C57BL/6 mice, 9 weeks old, were randomized into myocardial infarction group, embryonic stem cells group, H2O2 pretreatment group, with eight mice in each group, which were respectively injected 150 μL PBS, 150 μL embryonic stem cells (1×106), and 150 μL H2O2-pretreated embryonic stem cells (1×106) at 1 week after modeling. At 3 weeks after transplantation, left ventricular end-diastolic diameter, left ventricular end-systolic inner diameter and left ventricular ejection fraction of mice in each group were measured by ultrasound. Myocardial collagen content was detected by Sirius red staining and the average collagen volume fraction was calculated. RESULTS AND CONCLUSION: Compared with the myocardial infarction group, the left ventricular end-diastolic diameter and left ventricular end-systolic inner diameter were significantly decreased in the embryonic stem cells  group and H2O2 pretreatment group, while the left ventricular ejection fraction was significantly increased (P < 0.05). Heart weight/body weight, left ventricular weight/body weight, and average collagen volume fraction were significantly lower in the embryonic stem cells group and H2O2 pretreatment group than the myocardial infarction group (P < 0.05). Therefore, in vitro H2O2 pretreatment enhances reduce the post-myocardial infarction myocardial fibrosis and greatly improve heart 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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Thyroxine effects on induced differentiation of human umbilical cord blood-derived mesenchymal stem cells into chondrocytes Yang Tao, Jiang Bo, Xu Peng, Lin Gang 2015, 19 (1):  7-11.  doi: 10.3969/j.issn.2095-4344.2015.01.002 Abstract ( 299 )   PDF (666KB) ( 599 )   Save BACKGROUND: Human umbilical cord blood-derived mesenchymal stem cells can be induced through the co-culture to differentiate into other cells, but how to get more seed cells for tissue engineering is one of the most difficult problems. OBJECTIVE: To investigate the role of the different concentrations of thyroxine in chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells by co-culture with rabbit chondrocytes. METHODS: Human umbilical cord blood-derived mesenchymal stem cells were co-cultured with rabbit chondrocytes at 2:1, and stimulated by medium containing different concentrations of thyroxine (0, 0.01, 0.1 and 1, 10 μmol/L). Co-cultured cells with no thyroxine served as control group. After 14 days of co-culture, the cell RNA and protein were extracted, mRNA expressions of aggrecan and collagen type II were detected by real-time PCR, and protein expression of aggrecan and collagen type II were detected by western blot assay. RESULTS AND CONCLUSION: After intervention with 0.01, 0.1, 1, 10 μmol/L thyroxine, the mRNA and protein expressions of aggrecan and collagen type II were enhanced with the increase of thyroxine concentration, which were significantly different from those in the control group (P < 0.05). Experimental findings indicate that high levels of thyroxine can enhance the chondrogenic ability of human umbilical cord blood-derived mesenchymal stem cells co-cultured with rabbit chondrocytes. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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T-cell immune tolerance of HLA haploidentical donor induced by CTLA4Ig-modified bone marrow stromal cells Wang Ji-gang, Zhou Fan, Liu Yan-qin, Bai Ying, Liu Jing-hua, Zhang Hai-ting, Li Min-yan 2015, 19 (1):  12-17.  doi: 10.3969/j.issn.2095-4344.2015.01.003 Abstract ( 289 )   PDF (3035KB) ( 674 )   Save BACKGROUND: CTLA-4Ig as a tolerance-induction agent is a potential strategy in graft-versus-host disease prevention. OBJECTIVE: To investigate the efficacy of CTLA4Ig-gene-modified bone marrow stromal cells mediated by adenovirus to induce T-cell tolerance of haploidentical donors. METHODS: The bone marrow stromal cells isolated culture from the bone marrow of HLA haploidentical donors were transfected by recombinant adenovirus encoding CTLA4IgcDNA (AdCTLA4Ig) at a multiplicity of infection=50 for 72 hours. The expression rate and the location of CTLA4Ig in the transfected cells were detected by fluorescence microscope after immunofluorescence staining. CTLA4Ig-modified bone marrow stromal cells (2×104, 4×104 and 8×104) were respectively co-cultured with 105 T cells from the peripheral blood of HLA haploidentical donors and 105 peripheral blood mononuclear cells from recipients. The proliferative inhibition rate was determined by MTT assay, and the level of interleukin-2 in the supernatant was detected by ELISA. The bone marrow mononuclear cells (1×105/well) were co-cultured with CTLA4Ig-modified bone marrow stromal cell layers  constructed in 6-well plates. The number of bone marrow mononuclear cells and colony-forming unit-granulocyte macrophages were calculated after 5-day culture. RESULTS AND CONCLUSION: The expression rate of CTLA4Ig at the multiplicity of infection=50 was as high as 85%, and the immunofluorescence signals of CTLA4Ig were distributed unevenly in the cytoplasm. The inhibition rates of 2×104, 4×104, and 8×104 CTLA4Ig-modified bone marrow stromal cells on proliferation of T cells were higher than that of untransfected cells. The levels of interluekin-2 in the corresponding cell groups were significantly lower than that in the untransfected cells (P < 0.05). At 5 days of culture, there was no significant difference in the number of bone marrow mononuclear cells and colony-forming unit-granulocyte macrophages between the transfected and untransfected cell groups (P > 0.05). These findings indicate that CTLA4Ig-modified bone marrow stromal cells mediated by adenovirus can induce immune tolerance of T-lymphocyte from HLA haploidentical donors 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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Age-associated proliferation and differentiation changes of rat bone marrow mesenchymal stem cells Li Hai-rui, Zheng Dong, Jiang Can, Guo Jun, Zhang Ai-dong, Li Zi-cheng 2015, 19 (1):  18-23.  doi: 10.3969/j.issn.2095-4344.2015.01.004 Abstract ( 327 )   PDF (1842KB) ( 671 )   Save BACKGROUND: Autologous bone marrow mesenchymal stem cell transplantation for the treatment of cardiovascular disease has become one of the hotspots, but it is unclear whether the proliferation and directional differentiation of autologous bone marrow mesenchymal stem cells varies changes with age. OBJECTIVE: To explore the proliferation and differentiation changes of rat bone mesenchymal stem cells in different ages. METHODS: The bone marrow mesenchymal stem cells from Sprague-Dawley rats in different age groups were purified and cultured, and then examined by flow cytometry in terms of cell cycle. Meanwhile, neonatal rat cardiomyocytes were co-cultured with bone marrow mesenchymal stem cells. The morphologic changes of bone marrow mesenchymal stem cells and the protein expression of troponin T were detected with immunofluorescence technique. RESULTS AND CONCLUSION: The percentage of bone marrow mesenchymal stem cells in G0/G1 phase was increased with age; while the percentage of expression of troponin T proteins-positive bone marrow mesenchymal stem cells were decreased with age. These findings indicate that the proliferation and differentiation abilities of rat bone marrow mesenchymal stem cells descend with age. 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 culture method induces the chondrogenic differentiation of human adipose-derived mesenchymal stem cell microspheres Bi Xiao-juan, Guo Chen-ming, Li Liang, Duan Xian-lin, Jiang Ming 2015, 19 (1):  24-29.  doi: 10.3969/j.issn.2095-4344.2015.01.005 Abstract ( 422 )   PDF (2201KB) ( 1109 )   Save BACKGROUND: Adipose-derived mesenchymal stem cells are more appropriate as seed cells for cartilage induction in patients who have unsatisfied repair after articular cartilage injuries, but how to induce the cartilage function need to be studied. OBJECTIVE: To induce the chondrogenic differentiation of human adipose-derived mesenchymal stem cell microspheres using three-dimensional culture system. METHODS: Adipose tissues were extracted after liposuction under sterile, to isolate and culture human adipose-derived mesenchymal stem cells. The third passage cells were analyzed by flow cytometry and identified by osteogenic induction and adipogenic induction, and meanwhile, cells were cultured under three-dimensional culture system for chondrogenic differentiation. Then, cells were observed by Alcian blue staining for identification of glycosaminoglycan synthesis as well as hematoxylin-eosin staining for histological analysis. Expression of collagen type II was detected by immunofluorescence method. Cartilage stiffness was determined by mass measurement. RESULTS AND CONCLUSION: Isolated human adipose-derived mesenchymal stem cells highly expressed CD105, CD44, CD29, but showed low expressions of CD45, CD34. After osteogenic and adipogenic induction, cells were positive for alizarin red staining and oil red O staining. Under the three-dimensional culture, chondrocytes could greatly express glycosaminoglycans and collagen type II. The results suggest that under the three-dimensional culture, human adipose-derived mesenchymal cells differentiate into chondrocytes and have the characteristics of chondrocytes. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cells combined with peptide hydrogel and chondrogenic factors for repair of articular cartilage defects in rabbits Wang Yan, Li De-hua 2015, 19 (1):  30-36.  doi: 10.3969/j.issn.2095-4344.2015.01.006 Abstract ( 386 )   PDF (967KB) ( 559 )   Save BACKGROUND: Peptide hydrogel has good plasticity, and it can fill the injured site very well; therefore, to use this material as a scaffold is a feasible exploration in bone and cartilage tissue engineering. OBJECTIVE: To test the effect of bone marrow mesenchymal stem cells combined with injectable peptide hydrogel and chondrogenic factors for repair of articular cartilage defects in rabbits. METHODS: Bone marrow mesenchymal stem cells from rabbits were isolated and cultured. A full-thickness bone-cartilage defect model, 5 mm in diameter and 3 mm in depth, was made on the left knee joint of rabbits, and the right knee joint of rabbits with no treatment was used as control after modeling. There were three experimental groups: self-assembling peptide hydrogel group, peptide hydrogel+chondrogenic factors group, and peptide hydrogel+chondrogenic factors+bone marrow mesenchymal stem cells group. Transforming growth factor β1, dexamethasone and insulin-like growth factor 1 were mixed as chondrogenic factors and added into self-assembling peptide hydrogel or bone marrow mesenchymal stem cells. Twelve weeks after treatment, animals were sacrificed for gross and histological observation, X-ray radiography, and histological evaluation using immunohistochemistry method. RESULTS AND CONCLUSION: At 12 weeks after treatment, the self-assembling peptide hydrogel group showed excellent results in the cartilage repair, and better achievements in safranin-O staining, collagen II immunostaining, and histological scores than the other groups (P < 0.05); the peptide hydrogel+chondrogenic factors group had better repair effects similar to the self-assembling peptide hydrogel group, but the expression of proteoglycans was higher than that in the control group (P < 0.01); there were poorer repair effects and more osteophytes in the peptide hydrogel+chondrogenic factors+bone marrow mesenchymal stem cells group than the the peptide hydrogel+ chondrogenic factors group. Experimental findings indicate that the self-assembling peptide hydrogel can repair cartilage defects in situ and improve cartilage repair, which is expected to improve current repairing effects on cartilage defects. 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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Platelet-derived growth factor promotes skin wound healing by regulating the migration of bone marrow mesenchymal stem cells to wounds Ding Yue, Xu Hai-long, Xie Hong 2015, 19 (1):  37-43.  doi: 10.3969/j.issn.2095-4344.2015.01.007 Abstract ( 362 )   PDF (1030KB) ( 467 )   Save BACKGROUND: Platelet-derived growth factor has the ability of wound repair, and relevant studies mainly focus on bone tissue repair. However, there are less studies about the effect of platelet-derived growth factors in skin wound healing. OBJECTIVE: To investigate the role of platelet-derived growth factor to promote wound healing by the regulation of bone marrow mesenchymal stem cells to the wound. METHODS: Bone marrow mesenchymal stem cells from rats were cultured. Immunofluorescence method was conducted to detect cell surface markers of CD34 and CD44 in bone marrow mesenchymal stem cells. Thirty healthy male SD rats were divided into five groups at random. Bone marrow mesenchymal stem cells labeled with PKH-26 were injected into the rat caudal vein in each group. The rats were anesthetized 1 week after injection. On the center of rat back, a 3-cm incision was made to establish a wound healing model. Different concentrations of platelet-derived growth factor were injected via multi-points on the skin wound after modeling, and the control group was treated with the same volume of normal saline. Skin wound tissues were taken for relevant parameter measurement at 14 days after injection. RESULTS AND CONCLUSION: Under the fluorescence microscope, platelet-derived growth factor could induce the migration and accumulation of bone marrow mesenchymal stem cells to the trauma in a dose-dependent manner and promote the wound healing. Masson staining showed that, with the concentration increase, platelet-derived growth factors could reduce inflammatory cell infiltration and increase the number of collagen fibers. Results from western blot assay showed that platelet-derived growth factor could inhibit the expression of matrix metalloproteinase-1, promote the expression of tissue inhibitor of matrix metalloproteinase 1 in the wound, and inhibit the collagen degradation, thereby promoting skin wound healing indirectly. 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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Fluorescent supermagnetic nanoparticles-labeled adipose-derived mesenchymal stem cells in the three-dimensional culture system  Lv Pin-lei, Su Yue-han, Cui Da-xiang, Wang Zheng 2015, 19 (1):  44-48.  doi: 10.3969/j.issn.2095-4344.2015.01.008 Abstract ( 369 )   PDF (2107KB) ( 426 )   Save BACKGROUND: Fluorescent magnetic nanoparticles with the properties of quantum dots and magnetic particles have good biocompatibility, and can label cells effectively through endocytosis. OBJECTIVE: To validate the feasibility of fluorescent magnetic nanoparticles in labeling human adipose-derived mesenchymal stem cells. METHODS: Healthy human adipose tissue was extracted and adipose-derived mesenchymal stem cells were isolated in vitro by type I collagenase digestion. Passage 6 cells were incubated with the fluorescent magnetic nanoparticles overnight. Prussian blue staining and laser scanning confocal microscope were used to observe labeled adipose-derived mesenchymal stem cells in vitro after co-culturing with fluorescent magnetic nanoparticles. The tracing effect of labeled adipose-derived mesenchymal stem cells in vivo was detected by fluorescence imaging system. RESULTS AND CONCLUSION: Prussian blue staining showed that the fluorescent magnetic nanoparticles dispersed in the cytoplasm of adipose-derived mesenchymal stem cells in the form of blue particles. Under the laser scanning confocal microscope, the nuclei of adipose-derived mesenchymal stem cells were dyed blue by Hoechest33258, and the cytoplasm was dyed green. The fluorescence imaging results showed that labeled human adipose-derived mesenchymal stem cells had good imaging results. Therefore, as an efficient tracer, the fluorescent magnetic nanoparticles can label human adipose-derived stem cells in vitro and provide a new method for transplantation and transformation of adipose-derived 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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The expression of Wnt/beta-catenin signaling molecule in inflammatory bowel diseases treated with bone marrow mesenchymal stem cell transplantation Xing Yan-fen, Xie Xu-hong, Yuan Zhao-hu, Cui Ye-jia, Li Yu-yuan, Nie Yu-qiang, Wei Ya-ming 2015, 19 (1):  49-53.  doi: 10.3969/j.issn.2095-4344.2015.01.009 Abstract ( 233 )   PDF (1878KB) ( 406 )   Save BACKGROUND: The Wnt/β-catenin signaling pathway is one of the most important signaling pathways in stem cell regulation, which is involved in regulation of cell proliferation and differentiation. OBJECTIVE: To investigate the expression of Wnt/β-catenin main signaling molecule in inflammatory bowel tissues treated with bone marrow mesenchymal stem cell transplantation. METHODS: 2,4,6-Trinitrobenzene sulfonic acid was used for establishing inflammatory bowel diseases rat models. Bone marrow mesenchymal stem cells labeled with green fluorescent protein were transplanted into rat models via tail vein. Normal saline was injected as control. The expression of Wnt/β-catenin signaling molecule was detected in the large intestine tissue of inflammatory bowel disease rat models by quantitative RT-PCR at 14 and 28 days after transplantation. RESULTS AND CONCLUSION: Real-time quantitative PCR results showed that the expression of Wnt3a and  β-catenin in the inflammatory bowel tissue increased significantly (P < 0.05), while no difference in the expression of c-myc (P > 0.05). The expressions of Wnt3a, β-catenin and c-myc in the transplantation group were significantly lower than those in the control group after transplantation (P < 0.05). These findings indicate that the Wnt/β-catenin signaling pathway plays important roles in inflammatory bowel disease and repair after bone marrow mesenchymal stem cell transplantation, while this pathway may promote stem cells differentiating into intestinal epithelium, promote recovery from inflammatory bowel disease, repair inflammatory area, and restore intestinal tissue homeostasis. 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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Adipose-derived stem cells transfected with adenovirus carrying bone morphogenetic protein 14 for repair of articular cartilage injury Ma Hong-bin, Li Yun-xiang, Wang Ming-lun 2015, 19 (1):  54-60.  doi: 10.3969/j.issn.2095-4344.2015.01.010 Abstract ( 331 )   PDF (3131KB) ( 801 )   Save BACKGROUND: The articular cartilage has weak self-repair ability, mainly due to its lack of trophoblast cells in blood vessels and slow cell metabolism. Current treatment methods cannot restore the original function of the cartilage tissue, and cartilage tissue engineering in recent years has garnered increasing attention. OBJECTIVE: To observe the effect of adipose-derived stem cells transfected with bone morphogenetic protein 14 combined with type I collagen sponge scaffold on the repair of articular cartilage injury in the knee of rabbits. METHODS: Adipose-derived stem cells were isolated and cultured from rabbit subcutaneous adipose tissue, and transfected with Ad-CMV-BMP-14-IRES-hrGFP-1. Type I collagen sponge scaffold with the transfected adipose-derived stem cells was used to repair articular cartilage injury in the knee of rabbits. Twelve weeks after operation, the articular tissue was taken for gross assessment and histological evaluation. RESULTS AND CONCLUSION: The expressions of bone morphogenetic protein 14, type II collagen and Sox-9 were higher in cells transfected with bone morphogenetic protein 14 than untransfected ones. At 12 weeks after operation, adipose-derived stem cells transfected with bone morphogenetic protein 14 combined with type I collagen sponge scaffold had good repair effect on articular cartilage injuries, and the injured cartilage tissues were smooth and had good texture, color and integration junction; adipose-derived stem cells combined with type I collagen sponge scaffold could partially repair the injured cartilage tissues that had similar color and texture to normal tissues, and there was a remarkable boundary between the repaired tissue and normal cartilage tissue;  simple type I collagen sponge scaffold was almost collapsed, and no hyaline cartilage tissue formed. These findings indicate that transfection of bone morphogenetic protein 14 can strengthen the ability of adipose-derived stem cells dramatically to repair cartilage injuries. 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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Expression of nerve growth factor and neurotrophin 3 after transplantation of human umbilical cord blood stem cells combined with electroacupuncture stimulation in rats with spinal cord injuries Sun Zhao-zhong, Li Rui, Fang Qing-min, Wang Guang-lin, Geng Xiao-peng, Ren Jia-bin, Yang Cheng 2015, 19 (1):  61-66.  doi: 10.3969/j.issn.2095-4344.2015.01.011 Abstract ( 296 )   PDF (2277KB) ( 557 )   Save BACKGROUND: Studies have shown that umbilical cord blood stem cell transplantation promote the recovery of spinal cord injury, and electroacupuncture also can inhibit the proliferation of astrocytes to reduce damage to scar formation, suggesting that a combination of umbilical cord blood stem cell transplantation and electroacupuncture may play an important role in the treatment of acute spinal cord injuries. OBJECTIVE: To observe the influence of transplantation of human umbilical cord blood stem cells combined with electroacupuncture at the Du channel on expression of nerve growth factor and neurotrophin 3 in rats with spinal cord injuries. METHODS: Seventy-two female Sprague-Dawlay rats were randomly divided into control group, injury group, transplantation group and combined therapy group. In the control group, only an incision on the back was sutured; in the injury group, a piece of saline-infiltrated gelatin sponge, 1 mm×2 mm×2 mm, was placed into the transected spinal cord at T10 level; in the transplantation group and combined therapy group, a piece of gelatin sponged infiltrated in the suspension of human umbilical cord blood stem cells was placed into the transected spinal cord, respectively, and then, electroacupuncture stimulation at the Du channel was performed in the combined therapy group at 1 hour after modeling. Specimens were taken at 7, 14, 28 days after modeling in each group, and then immunohistochemistry, western blot and real time-PCR methods were used to detect the expression of nerve growth factor and neurotrophin 3. RESULTS AND CONCLUSION: Compared with the transplantation group, the expression of nerve growth factor and neurotrophin 3 was lower in the injury group but higher in the combined therapy group at 7, 14, 28 days after modeling(P < 0.05). The results of western blot and real time-PCR were consistent with those of immunohistochemical detection. Findings show that human umbilical cord blood stem cell transplantation combined with electroacupuncture has a remarkable synergistic effect in the treatment of spinal cord injury that can significantly up-regulate the expression of nerve growth factor and neurotrophin 3, and contribute to injured spinal cord repair, regeneration and functional recovery after spinal cord 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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How does autophagy activation affect the apoptosis, proliferation and cycle of endothelial progenitor cells in rats? Liu Hui, Li Xiao-qiang, Zhu Ren-da, Meng Qing-you, Lu Hui-jun 2015, 19 (1):  67-71.  doi: 10.3969/j.issn.2095-4344.2015.01.012 Abstract ( 285 )   PDF (2021KB) ( 304 )   Save BACKGROUND: Previous studies have reported that rapamycin can affect the proliferation, migration and adhesion abilities of endothelial progenitor cells, but there is no report on the effect of autophagy, as well as the interaction between autophagy and apoptosis.   　 OBJECTIVE: To observe the effect of rapamycin activated autophagy activation on the proliferation, apoptosis, and cycle of endothelial progenitor cells.  METHODS: Density gradient centrifugation was used to obtain mononuclear cells from bone marrow, and the mononuclear cells were inoculated on human fibronectin-coated culture plate.Then after cultured for 7 days the adherent cells collected were the endothelial progenitor cells. Different concentrations of rapamycin (0.01, 0.1, 1 and 10 μg/L) were added and cultured for 24 hours. Western blot was used to detect the LC3-II protein expression and monitor the induction of autophagy, flow cytometry was used to observe the cell cycle progression and apoptosis changes, and methylthiazolyldiphenyl-tetrazolium bromide colorimetric assay was used to observe the proliferation ability. Meanwhile, the ultrastructural changes were observed under transmission electron microscope.  RESULTS AND CONCLUSION: Compared with the control group, there was no significant increasing of LC3-II protein expression of endothelial progenitor cells in 0.01 μg/L rapamycin group, and the LC3-II protein expression was in the high level. The LC3-II protein expression in the 1 μg/L and 10 μg/L rapamycin groups was higher than that in the control group, but lower than that in the 0.01 μg/L rapamycin group, which indicated that autophagy was particularly active when the concentration of rapamycin was 0.01 μg/L. The apoptosis of endothelial progenitor cells was increased with the increasing of concentration of rapamycin, and the proliferation rate was decreased with the increasing of concentration of rapamycin. The results indicate that activation of autophagy by bapamycin can promote the cell apoptosis, change the cell cycle significantly, and can inhibit the proliferation of endothelial 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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Ex-vivo expansion of autologous adipose-derived stem cells for the recovery of nasal mucosal function Liu Yang, Jia De-jin, Yan Jun-ling, Li Liang, Chen Chong, Wang Cheng, Ding Hong, Tang Su-yang 2015, 19 (1):  72-77.  doi: 10.3969/j.issn.2095-4344.2015.01.013 Abstract ( 368 )   PDF (2365KB) ( 1180 )   Save BACKGROUND: The ex-vivo expanded autologous adipose-derived stem cells have the capability of multipotential differentiation and have a broad application prospect in the field of tissue engineering and regenerative medicine. OBJECTIVE: To observe the nasal mucosal structural repair and functional reconstruction using ex-vivo expanded autologous adipose-derived stem cells. METHODS: Ten patients with mucosal damage due to the physical or chemical factors were enrolled, including six cases of mucosal scar and four cases of mucosal ulceration. Autologous adipose tissue was extracted for in vitro isolation, culture and expansion of adipose-derived stem cells. Before transplantation, quality safety testing was done. All the patients were injected adipose-derived stem cells (1×107/cm2) at an interval of 15 days, totally for three times. Nasal volume, minimum cross-sectional area, and mucociliary clearance function were determined at 30, 90, 150 days after the final injection. Three of 10 patients were selected to take a 0.1 cm×0.1 cm mucosal tissue sample at 30 days before and after transplantation for hematoxylin-eosin staining, Masson trichrome staining, and AB-PAS staining. RESULTS AND CONCLUSION: Clinical symptoms were alleviated in all patients undergoing transplantation of adipose-derived stem cells. Compared with the baseline data, the nasal volume and minimum cross-sectional area were both decreased at 30, 90, 150 days after transplantation (P < 0.05), and the mucociliary clearance function was improved but not significantly (P > 0.05). Compared with the baseline data, the inflammation of the nasal mucosa was significantly reduced, collagen fibers arranged neatly, the deposition was decreased, and mucin secreted from goblet cells was increased in the selected three patients at 30 days after cell transplantation. These findings indicate that ex-vivo expanded autologous adipose-derived stem cells can be used to reconstruct the nasal mucosal structure and its 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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Granulosa cells with stem cell properties in the rat ovary Liu Hui-ping, Lin Zheng-hua, Zhang Guo-min, Yu Rong, Liu Wen-e, Li Ling, Gu Xu-yu 2015, 19 (1):  78-84.  doi: 10.3969/j.issn.2095-4344.2015.01.014 Abstract ( 562 )   PDF (2244KB) ( 674 )   Save BACKGROUND: Human and rat ovarian granulosa cells in dominant follicles have the phenomenon of expressing stem cell characteristics. OBJECTIVE: To investigate the expression of stem cell-related factors in rat ovarian granulosa cells. METHODS: After the paraffin sections of rat ovarian tissue, immunohistochemical method was used to detect CD34, CD133, ABCG2/Bcrp1, Pou5f1/Oct-4 expressions. Granulosa cells cultured in vitro were harvested by follicular puncture method, and then the immunohistochemical method was used to detect the expression of FSHR receptor in order to identify the purity of granule cells. In the cultured granulosa cells, CD44 and C-Kit expressions were detected immunohistochemically, RT-PCR was used to detect ABCG2/Bcrp1, Pou5f1/Oct-4, Nanog gene expressions in ovarian tissue and granulosa cells. RESULTS AND CONCLUSION: Immunohistochemistry detection on paraffin sections showed that a part of ovarian granulosa cells expressed CD34, CD133, ABCG2/Bcrp1 and Pou5f1/Oct-4, and the expression of Pou5f1/Oct-4 protein gradually increased in the development of ovarian follicles, significantly enhanced during the luteal phase, and then disappeared after the formation of corpus albicans, displaying a periodic expression characteristics. FSHR receptor positive identification rate of primary cells harvested by the folliclar puncture method was more than 95%. Granulosa cells cultured in vitro were mainly long spindle-shaped or diamond, and some cells presented with aggregation growth and expressed CD44 and C-Kit. RT-PCR test results showed that there were no Nanog in the ovarian tissue and cultured granulosa cells, low expression of Pou5f1/Oct-4 in the ovarian tissue, strong expression of ABCG2/Bcrp1 in the ovarian tissue, weak expression of Pou5f1/Oct-4 in the cultured granulosa cells, and strong expression of ABCG2/Bcrp1 in the cultured granulosa cells. These findings suggest that a part of granulosa cells in the rat ovarian have the characteristics of 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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Effects of transfection with acidic fibroblast growth factor by electroporation on skeletal muscle satellite cells Li Jiang-hua, Dong Shao-hong, Xiong Wei, Pang Xin-li, Liu Qi-yun, Li Wen-jun 2015, 19 (1):  85-90.  doi: 10.3969/j.issn.2095-4344.2015.01.015 Abstract ( 295 )   PDF (1847KB) ( 471 )   Save BACKGROUND: Previous studies have shown that a certain dose of acidic fibroblast growth factor can promote skeletal muscle satellite cell proliferation in vitro. OBJECTIVE: To investigate the effects of transfection with acidic fibroblast growth factor by electroporation on growth, proliferation and differentiation of skeletal muscle satellite cells. METHODS: Skeletal muscle satellite cells were cultured and purified, and then transfected with plasmid pSectag-GFP-aFGF by electroporation. The expression of green fluorescent protein was observed under fluorescence microscope, and the transfection efficiency was calculated. After transfection, cell cycle was analyzed by flow cytometry to draw the growth curve of skeletal muscle satellite cells. Western blot assay was employed to measure protein level of acidic fibroblast growth factor. RESULTS AND CONCLUSION: (1) Immunocytochemistry detection: The skeletal muscle satellite cells were positive for a-sarcomeric actin. (2) Transfection efficiency: At 12 hours after transfection with pSectag-aFGF, several cells showed green fluorescence, and the green fluorescent expression reached the peak at 72-96 hours after transfection with a positive rate of about 90%. (3) Cell cycle: After electrotransfection, the proportion of cells at S phase in the electroporation group was higher than that in the control group (P < 0.05). (4) Cell growth curve: At 3 days after electrotransfection, the cells entered logarithmic growth phase but the proliferation slowed down at 5 days. (5) Differentiation capacity: There were fewer myotubes and aging cells in the electroporation group than the control group. (6) Western blot assay: Acidic fibroblast growth factor protein was highly expressed in the cells transfected with target gene detected by western blot assay. These findings indicate that by using electroporation method, acidic fibroblast growth factor can be transferred into skeletal muscle satellite cells and have a high-efficiency and long-term expression, which can promote the proliferation of skeletal muscle satellite cells and inhibit formation of myotubes. 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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Tissue explant method versus enzymatic digestion method for culture of rat hair follicle stem cells Li Jia, An Heng-qing, Wang Feng, Wang Wen-guang, Paluoke Dilimulati, Wang Yu-jie, Mulati Rexiati 2015, 19 (1):  91-95.  doi: 10.3969/j.issn.2095-4344.2015.01.016 Abstract ( 324 )   PDF (1960KB) ( 560 )   Save BACKGROUND: Hair follicle stem cells have been confirmed to have stronger proliferative ability than interfollicular epidermal stem cells, which have been an issue of concern in seed cell research. OBJECTIVE: To compare the biological characteristics of rat hair follicle stem cells cultured by tissue explant method and enzymatic digestion method. METHODS: Under stereomicroscope, hair follicles were isolated from the rat whiskers, and then tissue explant method and two-step enzymatic digestion method were employed to culture hair follicle stem cells. Cells were purified using repeated differential adhesion method, and cell growth and morphology were observed periodically. Flow cytometry was used to detect the expression of CD34 and β1 integrin in passage 3 hair follicle stem cells. RESULTS AND CONCLUSION: Cells cultured by two-step enzymatic digestion method grew faster with more amount than those cultured by tissue explants method. Flow cytometry showed that the expressions of PE-CD34 and FITC-β1 were (39.52±19.57)% and (93.46±4.73)% for the two-step enzymatic digestion group, and (19.20±11.53)% and (363.57±14.42)% for the tissue explant method, respectively. There was a significant difference between the two methods. In conclusion, these two methods are able to culture high-activity hair follicle stem cells, which can be chosen according to different experimental requirements. 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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Long-term stable culture of canine vaginal epithelial cells and smooth muscle cells in vitro Shen Fu-jin, Luo Ruo-yu, Liang Hua, Jiang Yan-ping, Cao Lai-ying 2015, 19 (1):  96-100.  doi: 10.3969/j.issn.2095-4344.2015.01.017 Abstract ( 305 )   PDF (1680KB) ( 372 )   Save BACKGROUND: In vitro culture of sufficient vaginal epithelial cells and smooth muscle cells is the key for vaginal tissue engineering. However, the culture, purification and passage of vaginal epithelial cells in vitro are difficult. Primary culture and passage of vaginal epithelial cells from large animals such as canines has not been reported. OBJECTIVE: To establish a stable method of culturing canine vaginal epithelial cells and smooth muscle cells. METHODS: Vaginal epithelial cells were isolated from the vaginal specimens by enzymatic digestion with Dispase and trypsin separately, and cultured in keratinocyte serum-free medium. Vaginal smooth muscle tissue were minced and digested with collagenase type II; the collected smooth muscle cells were cultured in DMEM culture medium containing 10% fetal bovine serum. The cultured cells were passaged regularly. Cell morphology and proliferation characteristics were observed and cell phenotypes were confirmed by morphology and immunohistochemistry staining. RESULTS AND CONCLUSION: Primary vaginal epithelial cells began to adhere after 24-36 hours, grew logarithmically after 4-5 days, and reached 70% confluence after 7-8 days; the epithelial cells showed a typical cobblestone, with no fibroblasts. Cultured epithelial cells passaged every 4-5 days and subcultured to 6-7 generations continuously. Immunohistochemical staining confirmed a positive staining for anti-pancytokeratin (AEl/AE3). Primary cultured smooth muscle cells adhered and grew after 24 hours. The smooth muscle cells were spindle-shaped and proliferated logarithmically. After 4 days, primary cultured smooth muscle cells were confluent and showed a typical shape of “peaks and valleys”, and then the cells could be passaged every 3-4 days and passaged 7-8 generations. Immunohistochemistry staining showed α-actin staining was positive. These findings indicate that canine vaginal epithelial cells and smooth muscle cells could have a long-term stable culture and proliferation, to provide adequate seed cells for vaginal 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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Effects of paeoniflorin on the proliferation of bone marrow mesenchymal stem cells Chao Er-tao, Bai Hai, Wang Cun-bang, Xi Rui, Ou Jian-feng, Zhao Qiang 2015, 19 (1):  101-107.  doi: 10.3969/j.issn.2095-4344.2015.01.018 Abstract ( 293 )   PDF (2247KB) ( 406 )   Save BACKGROUND: Studies have shown that paeoniflorin functions as replenishing blood and treatment of autoimmune diseases, and bone marrow mesenchymal stem cells also play an important role in the body’s blood and immune function. However, paeoniflorin effects on bone marrow mesenchymal stem cell proliferation and cytokine secretion and expression are rarely reported. OBJECTIVE: To investigate the effect of paeoniflorin on proliferation of bone marrow mesenchymal stem cells and the expression of interleukin-6. METHODS: Human bone marrow mesenchymal stem cells were separated and cultured in vitro by density gradient centrifugation combined with attachment method. The biological characteristics of bone marrow mesenchymal stem cells were identified by flow cytometry and osteogenic/adipogenic induction. The proliferation of bone marrow mesenchymal stem cells under different concentrations of paeoniflorin was detected by MTT method. The mRNA expression and secretion of interleukin-6 in the supernatant of bone marrow mesenchymal stem cells were detected by RT-PCR and ELISA, respectively. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells were isolated successfully and had osteogenic and adipogenic differentiation potential. Compared with the control group, 2 μmol/L and 10 μmol/L paeoniflorin significantly promoted the proliferation of bone marrow mesenchymal stem cells. 10 μmol/L paeoniflorin could significantly decrease the proportion of bone marrow mesenchymal stem cells in G0/G1 phase and increase this proportion in S phase. Compared with the control group, the experimental group could significantly increase the secretion and mRNA expression of interleukin-6 (P < 0.01). It is concluded that paeoniflorin at certain concentrations can obviously promote the proliferation of bone marrow mesenchymal stem cells, and increase the expression and secretion of interleukin-6. 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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Transplantation of bone marrow stromal stem cells into the ischemic myocardium reduces myocardial apoptosis but cannot improve cardiac function Jiang Shuai, Dong Shi-cai, Wei Dong-xing 2015, 19 (1):  108-113.  doi: 10.3969/j.issn.2095-4344.2015.01.019 Abstract ( 424 )   PDF (865KB) ( 491 )   Save BACKGROUND: Bone marrow stromal stem cells transplanted into infracted cardiac tissue can inhibit and reduce myocardial apoptosis, but whether this effect is correlated with improvement in cardiac function is still unclear. OBJECTIVE: To study the early effect of bone marrow stromal stem cells transplanted into the ischemic myocardium on the cardiac function. METHODS: Models of acute myocardial infarction were established by ligation of the left anterior descending branch, while no ligation was done in the sham group. In the transplantation group, rat bone marrow stromal stem cells (0.1 mL, 2×106) were injected into five sites on the edge of infarcted myocardial tissues at 30 minutes after myocardial infarction. In the sham group and model group, the same volume of normal saline was injected into the myocardial tissues. Three days after cell transplantation, hemodynamic monitoring, echocardiography, TUNEL assay were employed to detect myocardial apoptosis. RESULTS AND CONCLUSION: At 3 days after cell transplantation, myocardial apoptosis was more evident in the infarct and ischemic zones of the model group than the sham group; the number of apoptotic myocardial cells was significantly lower in the infarct and ischemic zones of the transplantation group than the model group. Compared with the sham group, the mean arterial blood pressure and left ventricular systolic pressure were significantly reduced, the left ventricular end diastolic pressure was increased, and the left ventricular ejection fraction and shortened fraction were significantly lowered in the model and transplantation groups (P < 0.05), but there was no difference between the model and transplantation groups (P > 0.05). These findings indicate that myocardial apoptosis can be reduced but the cardiac function cannot be improved in acute myocardial infarction rats at early stage after bone marrow stromal stem cells transplantation. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Autologous peripheral blood mononuclear cell transplantation with porous core decompression for treatment of avascular necrosis of the femoral head: an 11-month follow-up evaluation Wang De-feng, Zhang Xi-shan, Wu Jing-guo, Tao Ru 2015, 19 (1):  114-118.  doi: 10.3969/j.issn.2095-4344.2015.01.020 Abstract ( 298 )   PDF (796KB) ( 573 )   Save BACKGROUND: A large number of clinical trials have found that the number of bone marrow stem cells at the femoral neck and proximal femur in patients with osteonecrosis of the femoral head is significantly reduced, accompanied by decreased activity, which causes a significant decrease in osteogenic capacity that the necrotic bone cannot be effectively repaired after absorption, leading to the collapse of the femoral head. OBJECTIVE: To probe into the early clinical efficacy of autologous peripheral blood mononuclear cell transplantation with porous core decompression for treatment of avascular necrosis of the femoral head. METHODS: Forty-five patients with avascular necrosis of the femoral head (49 hips) were enrolled in this study, and underwent autologous peripheral blood mononuclear cell transplantation with porous core decompression. After treatment, pain scores, Harris hip score, scores on the satisfaction of patients were evaluated, as well as X-ray, CT and MRI examinations. RESULTS AND CONCLUSION: All the patients received a follow-up visit of 11-14 months, averagely (12.5±0.6) months. During the follow-up, there were no complications and serious adverse reactions. Postoperative pain scores and Harris scores were both improved significantly compared with preoperative ones (P < 0.05). At 12 months after treatment, the excellent satisfaction rate was up to 92%. Patient’s MRI low signal region accounting for a percentage of the volume of the femoral head was decreased from (40.1±7.34)% preoperatively to (20.23±5.4)% at 6 months postoperatively, and there was a significant difference (P < 0.05). These findings indicate that autologous peripheral blood mononuclear cell transplantation with porous core decompression for treatment of avascular necrosis of the femoral head has significantly clinical effects at early stage, which can obviously reduce joint pain, improve and restore hip joint function, and delay progression of disease.  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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Cyclic mechanical stretch influences cell adhesion and spreading of immortalized human keratinocytes Liu Kun, An Mei-wen, Wang Li, Huang Jing-jing 2015, 19 (1):  119-123.  doi: 10.3969/j.issn.2095-4344.2015.01.021 Abstract ( 340 )   PDF (674KB) ( 474 )   Save BACKGROUND: The mechanical environment of skin tissue and spreading state of epithelial cells are closely related with the wound healing and scar formation process. OBJECTIVE: To analyze the effect of extracellular mechanical stimulation on cell spreading, to test the cell proliferation in order to analyze the effect of spreading form on cell proliferation and other physiological activities. METHODS: Cyclic sine wave mechanical stretching was exerted on immortalized human keratinocyte by using FX-4000 flexible substrate loading system, on the condition of 0.2 Hz and at frequency of 10% amplitudes. The spreading form was compared at 0, 24 and 48 hours, the cell proliferation was analyzed with flow cytometry, and the distribution of vinculin was analyzed with immunofluorescence staining. RESULTS AND CONCLUSION: human keratinocyte would keep the spreading state and could induce more cell proliferation by 24 hours mechanical stretching stimulation. Conversely, after stimulated for 48 hours, the morphology of the human keratinocyte was significantly changed, and the number of human keratinocyte in the division stage was larger than that in the static control group; under tensile stress, the distribution of vinculin was transformed from the surrounding nucleus membrane area to the cell edge. The results indicate that proper mechanical stimulate can increase cell proliferation with keeping cell spreading and adhesion state; the stimulating time of continuous cyclic stretching is the major factor to determine cell spreading morphology and adhesion regions of immortalized human keratinocyte. 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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Ease effect of ginsenoside on different-intensity ionizing radiation damage to human hematopoietic stem cells Huang Ying, Liang Xiao-yan, Li Cheng-jin, Hu Jun, Zhou Li-qian 2015, 19 (1):  124-129.  doi: 10.3969/j.issn.2095-4344.2015.01.022 Abstract ( 347 )   PDF (777KB) ( 487 )   Save BACKGROUND: Many domestic and foreign scholars and institutions are studying how to relieve radiation damage and to find the most suitable drug, while ginsenosides as the main pharmacological ingredient of ginseng show significant antioxidant effect. OBJECTIVE: To investigate the ease effect of ginsenosides on human hematopoietic stem cells under different intensities of ionizing radiations. METHODS: The CD34+ hematopoietic stem cells were isolated from the healthy cord blood. Then the cells were divided into normal group and ginsenoside-pretreated group, respectively, exposed under 1, 2, 5 Gy of X-ray irradiations for 24 hours. Cell viability was detected in irradiated hematopoietic stem cells by MTT assay. Apoptosis was estimated using the following assays: Annexin-V assay, caspase-3 mRNA and protein levels. The generation of reactive oxygen species was evaluated, in the presence or absence of ginsenoside in liquid cultures of CD34+ human hematopoietic stem cells irradiated with 1-, 2- and 5-Gy X-rays, using a flow cytometry assay. The Nrf-2 mRNA and protein levels were also studied by western blot analysis and RT-PCR, respectively. RESULTS AND CONCLUSION: Ionizing radiation at the therapeutic dose could decrease the viability of CD34+ cells and induce the cell apoptosis, and meanwhile, the activity of intracellular reactive oxygen species also showed a progressive increase that was correlated with the dose of ionizing radiation. However, ginsenoside pretreatment could relieve these above-mentioned effects. Ginsenoside inhibited the increase in caspase-3 activity induced by ionizing radiation, and additionally, enhanced the mRNA and protein expressions of Nrf-2 in CD34+ cells. In conclusion, ginsenoside protects CD34+ hematopoietic stem cells from radiation effects, which is probably correlated with anti-apoptosis and anti-oxidant roles of ginsenosides. 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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Expression of osteoblast-specific genes in immortalized rat dental follicle cells Zheng Hong 2015, 19 (1):  130-134.  doi: 10.3969/j.issn.2095-4344.2015.01.023 Abstract ( 242 )   PDF (692KB) ( 416 )   Save BACKGROUND: During in vitro culture, dental follicle cells are easy to loss self-renewal capacity and become aging, and they are also very difficult to be purified and amplified in quantity, which limits the application of dental follicle cells in periodontal tissue engineering. OBJECTIVE: To study the osteogenic effect of immortalized rat dental follicle cells. METHODS: pSSR69-pAmpho plasmids containing SV40T-Ag were used to transfect 293 cells. Rat dental follicle cells were transfected with virus supernatant and screened by hygromycin to establish immortalized rat dental follicle cells (experimental group). Untransfected cells served as controls. RT-PCR was used to detect the osteogenic related factors (alkaline phosphatase, osteocalcin, bone morphogenetic protein 2, and Runx2) in the experimental group and control group. RESULTS AND CONCLUSION: The results showed no statistical differences between the experimental group and the control group in the expressions of alkaline phosphatase, bone morphogenetic protein 2 and Runx2 (P > 0.05). The expression of osteocalcin was significantly higher in the experimental group than the control group (P < 0.05). These findings suggest that in the late osteogenesis differentiation, immortalized rat dental follicle cells may promote the secretion of osteocalcin and then make osteoblasts early entry into the bone calcification stage. 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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Identification of a novel human leucocyte antigen allele B*07:110 Zhou Yue, Li Jian-ping 2015, 19 (1):  135-139.  doi: 10.3969/j.issn.2095-4344.2015.01.024 Abstract ( 324 )   PDF (664KB) ( 549 )   Save BACKGROUND: In recent years, with the development of China Marrow Donor Program and the improvement of human leukocyte antigen (HLA) typing technique, novel alleles of human leukocyte antigen have been discovered constantly in China. OBJECTIVE: To identify and confirm a novel HLA allele in a Chinese individual. METHODS: A new HLA allele was found during routine human leukocyte antigen genotyping by PCR-sequence specific oligonucleotide probes and sequencing-based typing. RESULTS AND CONCLUSION: The HLA-B locus hybridization probe reaction patterns of this sample did not match with any known HLA-B alleles or allelic combinations. Exons 2, 3 were sequenced in both directions using HLA-B sequence primer and group-specific sequencing primer. The obtained sequence had 2nts change from B*07:02:01 at nt 226 and nt 228 where A->G (codon 76 ATA->GTG) and resulting in a coding change, 76 isoleucine (I) was changed to valine (V). This nucleotide sequence has been submitted to the GenBank nucleotide sequence database and it is available under the accession number HM989017. A novel HLA allele, HLA-B*07:110, was identified, and was named officially by the WHO Nomenclature Committee. 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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How to improve mesenchymal stem cells theropy for Duchenne muscular dystrophy? Zhu Hui, Pang Rong-qing, Pan Xing-hua, Ruan Guang-ping 2015, 19 (1):  140-145.  doi: 10.3969/j.issn.2095-4344.2015.01.025 Abstract ( 277 )   PDF (693KB) ( 604 )   Save BACKGROUND: Treatment of Duchenne muscular dystrophy is not ideal. In recent years, more and more scientists try to treat Duchenne muscular dystrophy with mesenchymal stem cells. OBJECTIVE: To summarize the mesenchymal stem cell treatment for Duchenne muscular dystrophy and guide relevant animal experiments in order to promote its clinical application. METHODS: A computer-based online search of CNKI and PubMed between January 2005 and June 2014 was performed to search related articles with the keywords of “Duchenne muscular dystrophy, mesenchymal stem cells, stem cell transplantation” in English and Chinese, respectively. Literatures related to mesenchymal stem cells for treatment of Duchenne muscular dystrophy were selected; in the same field, the articles published lately in authoritative journals were preferred. RESULTS AND CONCLUSION: A total of 317 literatures were primarily selected, and 41 articles were involved in result analysis according to inclusion criteria. Mesenchymal stem cells for treatment of Duchenne muscular dystrophy are mainly by increasing the number of homing cells and expression of dystrophin. Common methods are selecting the ideal seed cells, cell pretreatment, improving migration ways and intervening homing process. Mesenchymal stem cells treatment that has broad application prospects is an essential way for the treatment of Duchenne muscular dystrophy, and it can avoid the side effects of drug 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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Mechanism of umbilical cord mesenchymal stem cell therapy for metabolic syndrome Hu Shuang-shuang, Pan Xing-hua 2015, 19 (1):  146-150.  doi: 10.3969/j.issn.2095-4344.2015.01.026 Abstract ( 418 )   PDF (611KB) ( 450 )   Save BACKGROUND: Studies have shown that stem cells are directly involved in injury repair, secretion of growth factors to promote wound healing, improvement of blood circulation and promotion of angiogenesis, regulation of immune and inflammation, oxidative resistance, which may be used to treat a variety of chronic diseases. OBJECTIVE: To review the mechanism of umbilical cord mesenchymal stem cell therapy for metabolic syndrome. METHODS: A computer-based search of VIP, CNKI and PubMed was performed for articles concerning stem cell transplantation therapy for metabolic syndrome published from January 2004 to October 2014. The search terms were “stem cells, umbilical cord-derived mesenchymal stem cells, stem cells transplantation, metabolic syndrome, insulin resistance, diabetes, hyperlipidemia” in Chinese and English, respectively. Finally, 42 articles were included in result analysis. RESULTS AND CONCLUSION: Umbilical cord mesenchymal stem cells have the ability of self replication and differentiation, which are ideal seed cells for cell replacement therapy. Meanwhile, umbilical cord mesenchymal stem cells also have paracrine and immunomodulatory functions. Currently, technologies for isolation, culture, identification and in vitro amplification of umbilical cord mesenchymal stem cells are quite mature. Moreover, stem cell transplantation as a new method for the treatment of metabolic syndrome can fundamentally make the target organ damage better as well as improve insulin resistance and glucose and lipid metabolism disorder, which have been proved in animal experiments. However, there are still many clinical problems to be solved. 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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Expression of CD4+CD25+ regulatory T cells in aged patients with acute myeloid leukemia Bo Li-kui 2015, 19 (1):  151-155.  doi: 10.3969/j.issn.2095-4344.2015.01.027 Abstract ( 281 )   PDF (598KB) ( 631 )   Save BACKGROUND: High expression of CD4+CD25+ regulatory T cells is proved to exist in many patients with malignant tumors in vivo. Recent studies have found that CD4+CD25+ regulatory T cells also show a high percentage of expression in the peripheral blood of patients with acute myeloid leukemia. OBJECTIVE: To analyze the characteristics of expression of CD4+CD25+ regulatory T cells in aged patients with acute myeloid leukemia. METHODS: Ninety-two patients with newly diagnosed acute myeloid leukemia were enrolled and divided into middle-young age group (< 60 years old, n=22) and aged observation group (> 60 years old, n=70). Thirty-two patients in the aged observation group underwent standardized chemotherapy and showed complete remission, who acted as complete remission group; and the remaining 38 patients acted as aged group, including  6 cases of M2, 19 cases of M3, 7 cases of M4, and 6 cases of M5 according to FAB classification. At the same period, another 42 healthy persons for hospital examination were selected as control group. Peripheral blood samples were extracted to detect the expression of CD4+CD25+ regulatory T cells.  RESULTS AND CONCLUSION: The proportions of CD4+CD25high FOXP3+ Treg cells and CD4+FOXP3+ T cells in the aged and complete remission groups were higher than those in the control group (P < 0.01), and moreover, these two proportions were higher in the aged group than the complete remission group (P < 0.01). The proportions of CD4+CD25high FOXP3+ Treg cells and CD4+FOXP3+ T cells in the aged group were higher than those in the middle-young age group (P < 0.01). There were no difference in the proportions of CD4+CD25high FOXP3+ Treg cells and CD4+ FOXP3+T cells among different subgroups of the aged group (P > 0.05). The proportion of CD4+CD25high FOXP3+Treg cells in the peripheral blood was positively related to the proportion of CD4+FOXP3+ T cells in aged patients with acute myeloid leukemia (r=0.87, P=0.019). The proportion of CD4+CD25+ Treg cells in the aged patients with newly diagnosed acute myeloid leukemia is higher than that in healthy individuals and young patients with acute myeloid leukemia. 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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Instructions for Authors (2015)-Chinese Journal of Tissue Engineering Research (CJTER) 2015, 19 (1):  156-164.  doi: 10.3969/j.issn.2095-4344.2015.01.028 Abstract ( 411 )   PDF (701KB) ( 320 )   Save Chinese Journal of Tissue Engineering Research (CJTER, www.CRTER.org) is an open-access, peer-reviewed international journal. Depending on rapid publication, rigorous peer review process and professional editing services, CJTER strives to publish China’s latest progress in the exciting field of tissue engineering research, and to promote relevant academic exchanges as well. CJTER has been  awarded twice as “100 Outstanding Academic Journals of China”; and in 2013, the journal was titled “National Digital Publishing Demonstration Units” by the State Administration of Press, Publication, Radio, Film and Television of the People’s Republic of China. 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： Related Articles | Metrics