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02 July 2015, Volume 19 Issue 28 Previous Issue    Next Issue

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Bone marrow mesenchymal stem cells transfected with recombinant adenovirus vectors carrying basic fibroblast growth factor in co-culture with ligament fibroblasts Li Bin, Chen Liao-bin, Qi Yong-jian, Hu Dong-cai, Chen Biao 2015, 19 (28):  4429-4434.  doi: 10.3969/j.issn.2095-4344.2015.28.001 Abstract ( 313 )   PDF (1760KB) ( 373 )   Save BACKGROUND: Basic fibroblast growth factor (bFGF) plays an important role in the ligament tissue healing process, and bone marrow mesenchymal stem cells transfected with growth factors can be used as seed cells in ligament tissue engineering. OBJECTIVE: To observe biological effect of bone marrow mesenchymal stem cells transfected with recombinant adenovirus vectors carrying bFGF in three-dimensional co-culture with ligament fibroblasts. METHODS: Passage 3 bone marrow mesenchymal stem cells were divided into three groups: control group, Ad-EGFP group and Ad-bFGF group. Under a phase contrast microscope, the changes in cell morphology were observed and the rate of transfection was analyzed by flow cytometry. Proliferation of bone marrow mesenchymal stem cells and ligament fibroblasts was analyzed by MTS, the expression of bFGF protein in bone marrow mesenchymal stem cells was determined by ELISA. Scleraxis, collagen type I, collagen type III, decorin and cartilage oligomeric matrix protein levels were detected in BMSCs and ligament fibroblasts using real-time fluorescent quantitative PCR. RESULTS AND CONCLUSION: Recombinant adenovirus-mediated bFGF gene could transfect bone marrow mesenchymal stem cells efficiently. After co-culture for 3, 6 days, compared with the control group and Ad-EGFP group, in the Ad-bFGF group, the proliferation ability of bone marrow mesenchymal stem cells and ligament fibroblasts was enhanced (P < 0.01), the expression of bFGF protein in supernatant was obviously higher (P < 0.01), the collagen type I, collagen type III, decorin and cartilage oligomeric matrix protein mRNA expression decreased in the ligament fibroblasts (P < 0.01), but the mRNA expression of Scleraxis, collagen type I, collagen type III in bone marrow mesenchymal stem cells increased (P < 0.01). The results suggest that the co-culture of Ad-bFGF-transfected bone marrow mesenchymal stem cells with ligament fibroblasts promotes the proliferation of ligament fibroblasts while decreases the collagen synthesis at the same time, and stimulates the proliferation and differentiation of bone marrow mesenchymal stem cells into ligament fibrolasts.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Osteogenic induction of injectable geneX/bone marrow mesenchymal stem cells/ transforming growth factor beta 2 composite Lan Tian, Li Biao, Gong Yue-kun, Li Duo-yu, Bi Xin, Yang Yi 2015, 19 (28):  4435-4438.  doi: 10.3969/j.issn.2095-4344.2015.28.002 Abstract ( 316 )   PDF (776KB) ( 291 )   Save  BACKGROUND: Single scaffold materials have some shortcomings to some extent or in different ranges. Therefore, new composite materials are developed in recent year, which are compounded by two or more materials with complementary characteristics in a certain manner or ratio. OBJECTIVE: To investigate the histocompatibility and osteogenic induction of geneX artificial bone combined with bone marrow mesenchymal stem cells and transforming growth factor beta 2 (TGF-β2). METHODS: Injectable geneX was co-cultured with bone marrow mesenchymal stem cells for 5 days, and under the electron microscope, the histocompatibility of the artificial bone was observed. Passage 3 rabbit bone marrow mesenchymal stem cells were divided into three groups: simple cell group cultured with osteogenic medium, cell scaffold group cultured with geneX+osteogenic medium, combined group cultured with geneX+TGF-β2+ osteogenic medium. After 7, 14, 21 days, cell morphology, alkaline phosphatase activity detection, MTT detection, methyl thymol blue detection and alizarin red staining were performed. RESULTS AND CONCLUSION: After osteogenic induction, bone marrow mesenchymal stem cells were fibroblast-like cells and adhered to the surface of geneX bone with strong secretion of extracellular matrix. Cell proliferation and osteogenic activity in the combined group were stronger than those in the simple cell and cell scaffold groups (P < 0.05). The alkaline phosphatase activity was also higher in the combined group than the other two groups. These findings indicate that the geneX/bone marrow mesenchymal stem cells/TGF-β2 composite has good histocompatibility and pro-osteogenic differentiation ability.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | Related Articles | Metrics

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Effect of Longbie Capsule on proliferation of bone marrow mesenchymal stem cells Pan Jian-ke, Guo Bai-ming, Liu Jun, Zhang Xian, Xie Hui, Guo Da 2015, 19 (28):  4439-4444.  doi: 10.3969/j.issn.2095-4344.2015.28.003 Abstract ( 301 )   PDF (4397KB) ( 267 )   Save BACKGROUND:Longbie Capsule has satisfactory outcomes in the treatment of osteoarthritis, but its mechanism is still unclear. OBJECTIVE: To study the effect of Longbie Capsule on proliferation of bone marrow mesenchymal stem cells. METHODS: The bone marrow mesenchymal stem cells from SD rats were separated and expanded by whole adherence culture, then subcultured and confirmed by morphological observation and flow cytometry. Passage 4 cells were cultured in complete media containing 5 g/L, 1 g/L, 250 mg/L, 50 mg/L, 10 mg/L Longbie Capsule, respectively, for 24 hours. Then, MTT assay was used to detect cell viability. RESULTS AND CONCLUSION: The primary cells were adherent cells characterized by irregular shape, passage 2 cells were typically fibrous-shaped, passage 3 cells grew in long fibrous and swirl-type shape. Passage 4 cells were strongly positive for CD29 and CD90, positive for CD44, and negative for CD34 and CD45. 5 g/L and 1 g/L Longbie Capsule promoted the proliferation of bone marrow mesenchymal stem cells. These findings indicate that Longbie Capsule may promote the proliferation and differentiation of bone marrow mesenchymal stem cells, thereby playing a therapeutic effect on 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 Figures and Tables | Related Articles | Metrics

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Radix Astragali injection at appropriate concentrations enhances surface adhesion of bone marrow stromal cell Wang Cai-xia, Ren Ming-ji, Cui Ming-yu 2015, 19 (28):  4445-4449.  doi: 10.3969/j.issn.2095-4344.2015.28.004 Abstract ( 340 )   PDF (4749KB) ( 277 )   Save BACKGROUND: Several studies have demonstrated that many kinds of Chinese herbs for benefiting vital energy and enriching the blood secrete some cytokines by influencing bone marrow stromal cells to promote the differentiation and proliferation of hemopoietic stem cells or exert effects by promoting the adhesion of bone marrow stromal cells and hemopoietic stem cells. OBJECTIVE: To investigate the effects of Radix Astragali injection on the expression of intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 on the surface of mouse bone marrow stromal cells. METHODS: Mouse bone marrow stromal cells were isolated by adherent culture of whole bone marrow. The morphology of mouse bone marrow stromal cells were observed using inverted phase microscopy, hematoxylin-eosin staining, optical microscopy and transmission electron microscopy. The optimal concentration at which Radix Astragali injection exhibited the strongest effects on promoting the proliferation of bone marrow stromal cells was detected using MTT assay. Expression of intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 on the surface of mouse bone marrow stromal cells was detected using flow cytrometry after intervention by Radix Astragali injection. RESULTS AND CONCLUSION: Inverted phase microscopy and optical microscopy showed that bone marrow stromal cells adhered to the wall of culture flask, exhibited a shuttle-shaped or irregular appearance with processes. Through transmission electron microscopy, organelles were abundant, such as rough endoplasmic reticulum, secretory vesicle, and mitochondria. Radix Astragali injection at 400 and 600 mg/L significantly promoted the proliferation of mouse bone marrow stromal cells (P < 0.05) and there was no significant difference in the cell proliferation between these two concentrations. Radix Astragali injection at 600 mg/L increased the expression of intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 on the surface of mouse bone marrow stromal cells. These findings suggest that Radix Astragali injection at appropriate concentrations can increase the surface adhesion of bone marrow stromal cells, thereby improving hemopoietic microenvironment.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of curcumin on mesenchymalstem cells-induced immune tolerance to kidney transplantation Zhang Li-li, Lv Jun, Wan Xia, Xu An-ping 2015, 19 (28):  4450-4454.  doi: 10.3969/j.issn.2095-4344.2015.28.005 Abstract ( 534 )   PDF (5275KB) ( 353 )   Save BACKGROUND: Previous studies have shown that continuous administration of 1×107 of bone marrow mesenchymal stem cells can induce immune tolerance in rats undergoing kidney transplantation, but it is not yet found clinically that curcumin exerts an on immunomodulatory effect on kidney transplantation. OBJECTIVE: To investigate the effects of different concentrations of curcumin on bone marrow mesenchymal stem cells-induced immune tolerance in rats after kidney transplantation. METHODS: Rat model of kidney transplantation was made, and rat models were randomly divided into four groups: transplantation group with no treatment; bone marrow mesenchymal stem cells group (cell group) injected with 1×107/kg bone marrow mesenchymal stem cells via the left iliac vein (before peritoneal suture) and tail vein (from the 2nd day) for 10 days; bone marrow mesenchymal stem cells+low/high dosage of curcumin groups (low/high dosage curcumin groups) injected intragastrically with 2 or 10 mg/kg curcumin combined with injection of bone marrow mesenchymal stem cells for 10 days. Transforming growth factor-β1 protein expression in the kidney tissues was examined by immunohistochemical staining. The concentrations of interleukin-2 and interleukin-6 in serum were detected by ELISA assay. RESULTS AND CONCLUSION: After kidney transplantation, the protein expression of transforming growth factor-β1 in renal tubular epithelial cells and renal interstitial cells as well as the concentrations of interleukin-2 and interleukin-6 in serum were significantly higher in the transplantation group than the other groups (P < 0.05). Compared with the cell transplantation group, the protein expression of transforming growth factor-β1 as well as the concentrations of interleukin-2 and interleukin-6 reduced significantly in the low/high dosage curcumin groups (P < 0.05). These findings indicate that simultaneous administration of curcumin and bone marrow mesenchymal stem cells can effectively inhibit immune rejection reaction and improve renal function in rats after kidney transplantation. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Biocompatibility of bio-derived xenograft bone with bone marrow mesenchymal stem cells Gao Ming-jie, Tao Jie, Zhou Zi-hui, Du Lin 2015, 19 (28):  4455-4459.  doi: 10.3969/j.issn.2095-4344.2015.28.006 Abstract ( 344 )   PDF (947KB) ( 358 )   Save BACKGROUND: Bio-derived xenograft bone has natural pore structure of the bone, low immunogenicity, and good cytocompatibility. OBJECTIVE: To verify the biocompatibility of bio-derived xenograft bone with bone marrow mesenchymal stem cells. METHODS: Fresh pig femoral bone was collected to prepare bio-derived bone. Scanning electron microscope was used to observe the material structure. Passage 3 rabbit bone marrow mesenchymal stem cells at a density of 2×109/L were inoculated into the cancellous bone surface of the bio-dervied bone and cultured for 7 days. Cell growth was detected using scanning electron microscope. After culture for 8 days, cell number was counted. RESULTS AND CONCLUSION: The bio-derived bone had rough surface and irregularly interconnected pores constructing a mesh-like structure. After 3 days of compound culture, cells had irregular shapes and adhered to the surface of bio-derived bone; after 5 days of culture, cells were closely interconnected to form a layered growth; after 7 days of culture, cells exhibited multilayered growth and the extracellular matrix secreted locally. Under compound culture, the former 2 days were latent period, 3-6 days were logarithmic phase, and from the 6th day, the cell growth curve became smooth gradually and the cell proliferation decreased and entered into the plateau period. These findings indicate that the bio-derived xenograft bone has good biocompatibility with bone marrow mesenchymal stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Arthroscopic core decompression with autologous bone marrow induction material combined with titanium rod in the repair of stage II femoral head necrosis Wu Gang, Zhang Yong-feng 2015, 19 (28):  4460-4464.  doi: 10.3969/j.issn.2095-4344.2015.28.007 Abstract ( 343 )   PDF (814KB) ( 335 )   Save BACKGROUND: Although core decompression can contribute to the reconstruction of necrotic bone, its single use appears to have some deficiencies and cannot completely realize the reconstruction of the femoral head. OBJECTIVE: To explore the effect of arthroscopic core decompression with autologous bone marrow induction material combined with titanium rod in the repair of stage II femoral head necrosis METHODS: A retrospective analysis was performed on clinical data of 79 cases of stage II femoral head necrosis admitted at the South Branch of Tongchuan People’s Hospital from February 2011 to February 2013. According to the therapeutic methods, these cases were divided into control group (40 cases) and observation group (39 cases), and were given conventional core decompression and arthroscopic core decompression with autologous bone marrow induction material combined with titanium rod. Patients were followed up for 24 months, and the range of hip flexion and Harris scores on hip function and adverse events were observed and compared between two groups. RESULTS AND CONCLUSION: All the 79 patients successfully completed the 24-month follow-up. By the last follow-up, the range of hip flexion and Harris score were significantly higher in the observation group than the control group (P < 0.05). There was no failure case due to surgical material rejection. To sum up, arthroscopic core decompression with autologous bone marrow induction material combined with titanium rod is safe and effective for repair of stage II femoral head necrosis. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Migration and homing of bone marrow mesenchymal stem cells in segmental nerve injury Zhou Xue-feng, Ren Zhi-wu, Lu Ming, Wang Yu, Sun Zhen, Peng Jiang 2015, 19 (28):  4465-4471.  doi: 10.3969/j.issn.2095-4344.2015.28.008 Abstract ( 360 )   PDF (1211KB) ( 303 )   Save  BACKGROUND: A large number of studies have confirmed that tissue-engineered stem cell therapy is feasible to repair peripheral nerve injury, but the repair mechanism is unclear. OBJECTIVE: To observe the differentiation and homing of bone marrow mesechnymal stem cells under local nerve microenvironment by exploring the migration and effect of bone marrow mesenchymal stem cells in the repair of damaged nerve. METHODS: Male SD rats, aged 8 weeks, were selected to establish segmental nerve injury models by freezing the sciatic nerve. Thirty-six model rats were randomized into three groups (n=12): frozen nerve injury group, cell injection into the nerve group, cell injection around the nerve group. Before modeling and at 4, 8, 12 weeks after cell implantation, the sciatic nerve function index was measured. Electrophysiological test, contractility recovery rate, wet weight recovery rate of the triceps surae were detected and Masson staining was performed; toluidine blue staining of the distal nerve injury and immunofluorescence staining of the damaged nerve were performed. RESULTS AND CONCLUSION: At 4, 8, 12 weeks after cell implantation, the sciatic nerve function index was ranked as follows: frozen nerve injury group < cell injection around the nerve group < cell injection into the nerve group, but no significant difference was found among the three groups. Electrophysiological results showed that there was no difference in the latency and amplitude of compound muscle action potential between two cell therapy groups, but compared with the frozen nerve injury group, the latency was shorter and the amplitude was higher in the two cell therapy groups (P < 0.05). Cross-sectional area of the muscle fibers was lower in the frozen nerve injury group compared with the other two groups. Immunofluorescence staining showed that Nestin, S100 and P0 proteins were all expressed in the two cell therapy groups 12 weeks after cell implantation. These findings indicate that bone marrow mesenchymal stem cells added into the surrounding tissues of segmental damaged nerve can migrate into the damaged nerve, and even differentiate into Schwann cells and neural stem cells, which has a similar effect to the cell injection into the damaged cells.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Uric acid effect on Wnt signaling pathways during osteogenic differentiation of human bone marrow mesenchymal stem cells Wang Xiao-li, Xu Li-li, Yang Nai-long 2015, 19 (28):  4472-4477.  doi: 10.3969/j.issn.2095-4344.2015.28.009 Abstract ( 301 )   PDF (5002KB) ( 267 )   Save  BACKGROUND: Uric acid, as an endogenous antioxidant, exhibits anti-oxidative and anti-DNA damage effects, promotes osteogenic differentiation, and therefore has been paid more attentions. OBJECTIVE: To investigate the effects of different concentrations of uric acid on the expression of genes related to Wnt/β-catenin signaling pathways during osteogenic differentiation of human bone marrow mesenchymal stem cells.  METHODS: Healthy adult human bone marrow mesenchymal stem cells were in vitro isolatedby adherent culture of whole bone marrow. Passage 3 bone marrow mesenchymal stem cells were induced to differentiate into osteoblasts using different concentrations of uric acid (0, 140, 280, 560 μmol/L). Alkaline phosphatase activity, cell proliferation capacity, and Wnt-3α and β-catenin mRNA expression in the Wnt signaling pathways were detected. RESULTS AND CONCLUSION: Alkaline phosphatase staining was positive. After treatment with uric acid, alkaline phosphatase activity and cell proliferation capacity were increased, the expression of Wnt signaling pathway-related genes Wnt-3a and β-catenin was up-regulated in a dose-dependent manner. There were significant differences in the abovementioned indices between experimental groups and between each experimental group and control group (P < 0.05). These findings suggest that uric acid up-regulates Wnt-3a/β-catenin signaling pathway and thereby promotes the osteogenic differentiation and proliferation of human bone marrow mesenchymal stem cells in a dose-dependent manner.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Intravenous versus intratracheal administration of mesenchymal stem cells in a mouse model of asthma  Yao Yin, Guo Jie-bo, Deng Xue-quan, Sun Yue-qi, Fu Qing-ling 2015, 19 (28):  4478-4484.  doi: 10.3969/j.issn.2095-4344.2015.28.010 Abstract ( 338 )   PDF (6243KB) ( 316 )   Save BACKGROUND: Several studies have demonstrated that mesenchymal stem cells exhibit strong immunomodulation on allergic asthma. However, there are no reports to compare therapeutic effects under different administration ways. OBJECTIVE: To examine the immunomodulatory effects of MSCs via different administration routes on asthmatic mice. METHODS: Seventy-two Balb/c mice experienced three independent tests, and 24 mice were selected for each test. Twenty-four mice were randomly divided into four groups (n=6): control group, model group, intravenous treatment group and intratracheal treatment group. The mouse model of asthma was induced via intraperitoneal injection of ovalbumin at 1, 7, 14 days and 30-minute aerosol inhalation of ovalbumin at 22-26 days. In the latter two groups, mesenchymal stem cells were injected intravenously (200 μL, 5×109/L) or intratracheally (50 μL, 2×109/L) into the mice at 1 day before aerosol inhalation. RESULTS AND CONCLUSION: Compared with the control group, the airway inflammatory response was significantly increased in the model group. Intravenous administration of mesenchymal stem cells significantly alleviated the symptoms of allergic airway inflammation, including the airway hyperreactivity, the inflammatory cell counting in the bronchoalveolar lavage fluid, inflammatory cell infiltration in the lung tissue. Meanwhile, the levels of Th2 type cytokines in the bronchoalveolar lavage fluid and IgE in serum also decreased after intravenous administration of mesenchymal stem cells. However, the intratracheal application of mesenchymal stem cells did not exhibit the similar effects. Intravenous, not intratracheal, application of mesenchymal stem cells can exert immunomodulatory effects through the blood circulation.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Vascular endothelial growth factor 165 promotes proliferation of human adipose-derived mesenchymal stem cells  Wang Yu, Zhu Zhi-tu, Chen Jun-jiang 2015, 19 (28):  4485-4492.  doi: 10.3969/j.issn.2095-4344.2015.28.011 Abstract ( 306 )   PDF (1182KB) ( 514 )   Save BACKGROUND: Autologous fat transplantation has been widely used in soft tissue defect repair and cosmetic surgery, and the 1-year transplant survival rate is 20%-80%. Therefore, the establishment of timely and adequate blood supply at early period after transplantation is very important for the survival of transplanted fat tissues. OBJECTIVE: To observe the proliferation of human adipose-derived mesenchymal stem cells transfected with vascular endothelial growth factor 165. METHODS: Human adipose-derived mesenchymal stem cells were subcultured in vitro. Recombinant adenovirus carrying vascular endothelial growth factor 165 and blank virus were respectively transferred into adipose-derived mesenchymal stem cells. Cells cultured normally served as blank group. RESULTS AND CONCLUSION: Compared with the control and blank groups, the expressions of vascular endothelial growth factor 165 mRNA and protein were higher in the experimental group (P < 0.05). Experimental findings suggest that the recombinant adenovirus carrying vascular endothelial growth factor 165 transferred into adipose-derived mesenchymal stem cells cannot only maintain the expression of target protein but also obviously promote the proliferation 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 Figures and Tables | References | Related Articles | Metrics

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Osteogenesis of adipose stem cells on artificial bone scaffold  Su Peng, Zhang Wei-wei, Yu Hua-wei, Yang Han 2015, 19 (28):  4493-4497.  doi: 10.3969/j.issn.2095-4344.2015.28.012 Abstract ( 326 )   PDF (4938KB) ( 256 )   Save BACKGROUND: Insufficient source of seed cells is the important factor to restrict the tissue reconstruction and the development of regenerative medicine. OBJECTIVE: To explore the osteogenesis of adipose stem cells cultured with different kinds of artificial bones. METHODS: Adipose tissue was extracted from female volunteers undergoing cosmetic surgery to isolate adipose stem cells. Passage 4 adipose-derived mesenchymal stem cells were selected and divided into osteogenic induction group, osteogenic induction+hydroxyapatite group, osteogenic induction+absorbable bone group and osteogenic induction+recombinant bone xenograft group. The latter three groups were subdivided into 3, 10, 20 g/L subgroups, respectively. Culture medium was exchanged every 3 days, totally for 12 days. RESULTS AND CONCLUSION: Compared with the osteogenic induction group, the calcium concentration in the elution liquid was significantly higher in the osteogenic induction+hydroxyapatite group and low-concentration osteogenic induction+absorbable bone group (both P < 0.05), but no difference was found between the osteogenic induction group and high-concentration osteogenic induction+absorbable bone group (P < 0.05). In addition, the calcium concentration in the elution liquid was significantly lower in the osteogenic induction+ recombinant bone xenograft group than the osteogenic induction (P < 0.05). Therefore, different artificial bone scaffolds can influence the osteogenic effect of adipose stem cells, and among them, hydroxyapatite has a better effect on the osteogenic induction of adipose stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程   Figures and Tables | References | Related Articles | Metrics

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Extraction and identification of human adipose-derived stem cells  Wu Wei, Liang Fang, Song Xiao-qin, Hu Ping-an2 Liu Min 2015, 19 (28):  4498-4502.  doi: 10.3969/j.issn.2095-4344.2015.28.013 Abstract ( 1026 )   PDF (7207KB) ( 649 )   Save BACKGROUND: Adipose-derived stem cells are totipotent stem cells in the adipose tissue, and have the function of self-renewal and multi-directional differentiation. Human adipose-derived stem cells are ideal seed cells with stable genetic milieu and few rejections. OBJECTIVE: To extract human adipose-derived stem cells from human omental adipose tissue and to identify the cells by adipogenic and osteogenic induction. METHODS: Omental adipose tissues were collected from surgical patients to isolate and culture adipose-derived stem cells using type I collagenase digestion, filtration and centrifugation. Cell growth was observed and proliferative curve of human adipose-derived stem cells were drawn by cell counting method to calculate the doubling time at logarithmic growth phase. After adipogenic and osteogenic induction, induced cells were identified using oil red O and alizarin red staining, respectively. RESULTS AND CONCLUSION: Human adipose-derived stem cells were successfully isolated from the omentum tissues of surgical patients. Adherent cells were fusiform-shaped and like fibroblasts. The growth curve of passage 3 cells was in S shape, and the doubling time was 45.90 hours. After adipogenic and osteogenic induction for 2 and 3 hours, respectively, oil red O staining showed unequal-sized orange fat droplets, and alizarin red staining showed typical calcified nodules that were in orange. These findings indicate that adipose-derived . 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Different induction methods for bone marrow mesenchymal stem cells Dai Chuan-qiang1, Jia Xu-feng2, Zhang Lin3, Zhang Ge4 2015, 19 (28):  4503-4507.  doi: 10.3969/j.issn.2095-4344.2015.28.014 Abstract ( 267 )   PDF (907KB) ( 290 )   Save BACKGROUND: Transformation growth factor beta 1 is mostly used to induce the chondrogenic differentiation of bone marrow mesenchymal stem cells, but there is a poor induction efficacy. OBJECTIVE: To explore the chondrogenic differentiation of bone marrow mesenchymal stem cells co-cultured with articular chondrocytes or induced by transforming growth factor beta 1. METHODS: Articular chondrocytes and bone marrow mesenchymal stem cells from SD rats were harvested and divided into 1:2, 2:1, 1:1 concentration groups. Cells induced by transforming growth factor beta 1 acted as control group. After 20 days of induced culture, MTT was used to detect cell viability, alcian blue colorimetric assay was applied to measure glycosaminoglycan content, and western blot assay was employed to determine the expression of collagen type II. RESULTS AND CONCLUSION: The absorbance value in the control group was significantly lower than that in the 1:1 and 2:1 groups (P < 0.05). Glycosaminoglycan content and protein expression of collagen type II were also lower in the control group than the 1:2, 1:1, 2:1 groups. But there was no difference between 1:1 and 2:1 groups (P > 0.05). The results show that bone marrow mesenchymal stem cells co-cultured with articular chondrocytes can be induced to differentiate into chondrocytes, and meanwhile, there is a saturation phenomenon during the chondrogenic differentiation of bone marrow mesenchymal stem cells.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Biological characteristics of G666-1 nasopharyngeal cancer stem cells in Survivin gene silencing Wang Hui 2015, 19 (28):  4508-4513.  doi: 10.3969/j.issn.2095-4344.2015.28.015 Abstract ( 310 )   PDF (1090KB) ( 286 )   Save BACKGROUND: Survivin gene is the most potent anti-apoptosis factor, which can inhibit tumor cell proliferation and migration. It has become a hot topic related to cancer research in recent years. OBJECTIVE: To explore the effect of Survivin gene silencing on the biological characteristics of G666-1 nasopharyngeal carcinoma cells. METHODS: siRNA sequence of Survivin was designed and synthesized and then transfected into G666-1 cells using LipofectamineTM 2000. After Survivin gene silencing, Survivin mRNA and protein expressions were detected by RT-PCR and western blot assay, respectively. Expression of CD44+CD133+ cells was detected by flow cytometry. In addition, the effects of Survivin gene silencing on the proliferation, migration and invasion of G666-1 cells were evaluated through cell counting kit-8 test, scratch healing assay and cell invasion assay. RESULTS AND CONCLUSION: After Survivin gene silencing, the expressions of Survivin mRNA and protein were significantly reduced (P < 0.05); the growth of G666-1 cells was slowed dramatically, indicating the proliferation ability was inhibited, and the migration and invasion abilities were both decreased significantly (P < 0.05); the expression ratio of CD44+CD133+ cells was significantly lowered (P < 0.05). These findings indicate that the proliferation, migration and invasion of G666-1 nasopharyngeal cancer stem cells can be suppressed after Survivin gene silencing, and the proportion of CD44+CD133+ cells is reduced, thus indicating Survivin gene silencing may be a new effective target for treatment of nasopharyngeal carcinoma. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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KDR gene silencing effect on biological properties of breast cancer MCF-7 cells Yu Zhi-hao, Yu Yue, Yang Xin5, Cao Xu-chen 2015, 19 (28):  4514-4519.  doi: 10.3969/j.issn.2095-4344.2015.28.016 Abstract ( 372 )   PDF (1024KB) ( 308 )   Save BACKGROUND: With the development of genetic engineering and tumor molecular biology, gene therapy for tumors has become a new treatment modality. OBJECTIVE: To explore the effect of the KDR gene silencing on the proliferation and invasion capacity of breast cancer MCF-7 cells. METHODS: Interfering RNA (siRNA) sequences for small molecule KDR gene was designed and transferred into human breast cancer MCF-7 cells. Then, RT-PCR and western blot assay were used to detect the KDR mRNA and protein expression. Flow cytometry, cell counting kit-8 test and Transwell test were employed to detect the cell cycle, proliferative capacity and invasion capacity of breast cancer MCF-7 cells after the KDR gene silencing. RESULTS AND CONCLUSION: After 48 hours of KDR silencing, the mRNA and protein expressions of KDR in MCF-7 cells were decreased obviously; MCF-7 cells arrested at G0/G1 stage and the number of cells at S stage was reduced. Cell proliferation was inhibited significantly. The amount of cells passing through the filtering membrane became less. After KDR gene silencing, the proliferation and invasion of breast cancer MCF-7 cancer stem cells were inhibited remarkably, indicating that KDR gene silencing may be a new target for the effective treatment of breast cancer. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Allogeneic adipose-derived stem cells combined with demineralized bone matrix for repair of ulna defects: CT scan and histological detection Yang Han, Kang Jian-ping, Ding Yu-ming, Wang Song 2015, 19 (28):  4520-4525.  doi: 10.3969/j.issn.2095-4344.2015.28.017 Abstract ( 288 )   PDF (4393KB) ( 315 )   Save BACKGROUND: Adipose-derived stem cells have a wide variety of sources and strong proliferation ability, which are easy to access and simple to culture. Therefore, adipose-derived stem cells that are secondary to bone marrow mesenchymal stem cells are expected to become the most promising seed cells for bone tissue engineering. OBJECTIVE: To explore the feasibility of rabbit adipose-derived stem cells with demineralized bone matrix to repair ulna defects. METHODS: Ulna defect model was made in rabbits. Demineralized bone matrix was implanted into the right defect region as control group. After osteogenic induction, rabbit adipose-derived stem cells/demineralized bone matrix composite was implanted into the left defect region as experimental group. At 12 weeks after implantation, defect tissues were taken for CT scanning and histological detection. RESULTS AND CONCLUSION: CT results showed that there was unclear boundary between the broken ends of fractured bone and the composite material in the experimental group, and parallel calluses out of the broken end could be seen. In the control group, the broken end was clearly seen and no callus occurred continuously. Fibers were connected at the defect site, and no new bone occurred. Histological findings showed that typical regenerated bone tissues were seen in the experimental group with osteocytes, bone lacunae and bone trabeculae; there were more osteocytes and bone lacunae, but bone trabecula was only seen in a part of bone defects; a few of collagens interlarded the regenerated bone tissues. In the control group, the residual of demineralized bone matrix was seen as well as some collagenous fibers, and periosteal bone formed a little, but no large amount of regenerated osteoid tissues were found. These findings indicate that under osteogenic induction, rabbit adipose-derived stem cells combined with demineralized bone matrix are feasible to repair ulna 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 Figures and Tables | References | Related Articles | Metrics

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In vitro isolation, culture and identification of lung cancer stem cells in patients with lung squamous carcinoma Liu Zhe-liang1 Wu Jiao, Wang Lin-xian, Chen Yue-jun, Wu Guan-yu, Xiao Gao-ming 2015, 19 (28):  4526-4530.  doi: 10.3969/j.issn.2095-4344.2015.28.018 Abstract ( 360 )   PDF (5017KB) ( 336 )   Save BACKGROUND: Studies have shown that lung cancer stem cells can be isolated from lung cancer cell lines. But there are few reports about in vitro isolation, culture and identification of lung cancer stem cells in patients with lung squamous carcinoma. OBJECTIVE: To explore the feasible methods of harvesting lung cancer stem cells from fresh lung cancer tissue in patients with lung squamous carcinoma. METHODS: Side population cells were isolated by collagenase digestion, Ficoll density gradient centrifugation and Hoechst 33342 solution. The isolated cells were suspended in conditioned medium for isolated culture. Flow cytometry method was used to detect lung cancer stem cells based on the cell surface markers CD133 and CD44, and the positive rates of CD133+, CD44+ and CD133+/CD44+ cells were recorded. RESULTS AND CONCLUSION: Cells adhered at 0.5 hour after incubation; typical cell colony was formed at 4 days of culture; cells showed paving stone-shape at 7 days in a total number of 108. The positive rates of CD133+, CD44+ and CD133+/CD44+ cells at passage 4 were increased significantly. These findings indicate that stem   cell-like lung cancer cells were obtained from fresh lung cancer tissue in patients with lung squamous carcinoma, which were stably and rapidly amplified in vitro, laying the foundation for the further study on the heterogeneity and resistance of lung cancer stem cells in the future. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Bone marrow mononuclear cell transplantation for repair of traumatic brain injury via different approaches Zhao Nan, Liu Jun, Li Jun-yan, Ma Gang, Li Jin, Wang Ting-hua, Su Ping 2015, 19 (28):  4531-4536.  doi: 10.3969/j.issn.2095-4344.2015.28.019 Abstract ( 341 )   PDF (4708KB) ( 257 )   Save  BACKGROUND: There are several routes for stem cell transplantation; however, it is still unable to determine which one is the best. As for the different individuals with brain injury, the type of transplanted cells, transplantation route and time will affect the therapeutic effects. OBJECTIVE: To investigate the effect of bone marrow mononuclear cells transplanted via different approaches on neurological function of rats with traumatic brain injury. METHODS: Bone marrow mononuclear cells of rats were administered gradient centrifugation with Ficoll lymphocyte separation medium, and were labeled with CFDA-SE in vitro as standby. Rat models of traumatic brain injury were established by the method of freefall. After successful establishment of rat models, bone marrow mononuclear cells labeled with CFDA-SE were immediately transplanted into rats via injured area, lateral ventricle and internal carotid artery. One control group was designated for each transplantation route (bone marrow mononuclear cells were replaced with the same volume of DMEM). The degree of neurological deficits was evaluated using mNSS scores at different time points after treatment. The brain tissue was harvested after the last neurobehavioral evaluation. The survival and migration of bone marrow mononuclear cells in the injured area were observed under an inverted fluorescent microscope. RESULTS AND CONCLUSION: At 7, 10, and 14 days after treatment, the mNSS scores of rats in all groups were all lower than those at 1 and 3 days (P < 0.05). At 7 and 10 days, the mNSS scores of rats in the internal carotid artery transplantation group were significantly lower than those in the control group (P < 0.05). At 14 days after treatment, the number of fluorescence-labeled cells of rats in the internal carotid artery transplantation group was greater than that in the other groups (P < 0.05) and these labeled cells were widely distributed. The results demonstrate that the neurological function of rats can be improved by transplanting bone marrow mononuclear cells via the internal carotid artery, and a large number of transplanted cells can survive and migrate in the injured area.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Short-term efficacy of cardiac stem cells in improving the electrophysiological stability and ventricular fibrillation threshold after myocardial infarction  Zhong Ting-ting, Hou Jing-ying, Guo Tian-zhu, Zheng Shao-xin, Zhou Chang-qing, Long Hui-bao, Wu Quan-hua, Wu Hao, Wang Tong 2015, 19 (28):  4537-4543.  doi: 10.3969/j.issn.2095-4344.2015.28.020 Abstract ( 297 )   PDF (1172KB) ( 352 )   Save BACKGROUND: Our previous work demonstrated that cardiac stem cells (CSCs) transplantation could significantly improve the electrophysiological stability and ventricular fibrillation threshold in rats with myocardial infarction. OBJECTIVE: To compare the influence of CSCs and bone marrow mesenchymal stem cells (BMSCs) transplantation on the electrophysiological stability and ventricular fibrillation threshold in rats with myocardial infarction in the short term. METHODS: Thirty male healthy Sprague-Dawley rats were selected to make myocardial infarction models induced by the left anterior descending coronary artery ligation. Then, animals were randomly divided into three groups, CSCs group, BMSCs group and the PBS group, with 10 rats in each group. Two weeks after modeling, animals were respectively given the injection of 5×106 CSCs labeled with PKH26 in 0.1 mL PBS, 5×106 BMSCs labeled with PKH26 in 0.1 mL PBS or 0.1 mL PBS alone into the infracted anterior ventricular free wall. Two weeks after intervention, the electrophysiological characteristics and ventricular fibrillation threshold were measured respectively at the infarct zone, the infarct marginal zone and the non-infarct zone. Labeled CSCs and BMSCs were detected, and the expression of connexin-43 was examined in 5 µm cryostat sections from the infarct marginal zone of each heart. RESULTS AND CONCLUSION: Compared with the BMSCs group and PBS group, significant differences were revealed in the correct unipolar electrograms activation recovery time dispersion (ARTcd), electrical stimulation-induced malignant ventricular arrhythmias and ventricular fibrillation threshold at the infarct zone, the infarct marginal zone and the non-infarct zone in the CSCs group (P < 0.05). Obvious differences were discovered in the ARTcd, electrical stimulation-induced malignant ventricular arrhythmias and ventricular fibrillation threshold on the non-infarct area in the BMSCs group in contrast to the PBS group. Labeled CSCs or BMSCs were identified at the infarct marginal zone and expressed connexin-43. Connexin-43 was abundantly expressed in the CSCs group whereas it was rarely expressed in the BMSCs group, and even not expressed in the PBS group. These findings suggest that CSCs are superior to BMSCs in modulating the electrophysiological stability and the ventricular fibrillation threshold in the short term after transplantation, which is closely correlated with the expression of connexin-43.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Conditioned medium of bone marrow mesenchymal stem cells via intravenous injection to treat cerebral ischemia-reperfusion injury Chen Feng1, Liu Bin1, Ming Yuan-yuan, Zhou Su-qin, Shen Xia, Hua Fang, Cui Gui-yun, Yue Xuan-ye, Zan Kun, Ye Xin-chun 2015, 19 (28):  4544-4548.  doi: 10.3969/j.issn.2095-4344.2015.28.021 Abstract ( 245 )   PDF (910KB) ( 459 )   Save BACKGROUND: Large numbers of experimental data have confirmed that bone marrow mesenchymal stem cells have a positive therapeutic effect on cerebral ischemia-reperfusion injury, but there are few reports about intravenous administration of bone marrow mesenchymal stem cell conditioned medium in the treatment of stroke. OBJECTIVE: To investigate the effects of the conditioned medium of rat bone marrow mesenchymal stem cells on the recovery of neurological function in rats after cerebral ischemia-reperfusion injury. METHODS: The bone marrow mesenchymal stem cells were isolated from rat bone marrow. When cells at passage 2 or 3 reached 90% confluence, the original culture medium was removed. Then the cells were cultured in serum-free DMEM for 18 hours. After that, the culture solution was collected as the conditioned medium of rat bone marrow mesenchymal stem cells. Adult rats were subjected to 2 hours of right middle cerebral artery  occlusion. Ischemia-reperfusion injury rats were randomly assigned to three groups: control group, simple culture medium group and conditioned medium group, and respectively given injection of normal saline, DMEM, conditioned medium (10 mL/kg) via the tail vein at 2, 24, 48 hours after operation. RESULTS AND CONCLUSION: There was no difference in the behavioral tests among the three groups at postoperative 2 hours (P > 0.05). Compared with the control and simple culture medium group, neurological impairment was significantly improved in the conditioned medium group at postoperative 1, 3, 5 days (P < 0.05), but there was no significant difference between the control and simple culture medium groups. At postoperative 5 days, brain edema was significantly eased in the conditioned medium group in comparison to the control and simple culture medium groups (P < 0.05), and there was also no difference between the latter two groups (P > 0.05). These results suggest that rat bone marrow mesenchymal stem cells-conditioned medium via intravenous administration can significantly ease brain edema and improve the neurologic function after cerebral ischemia-reperfusion injury.    中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Telomerase reverse transcriptase genetic modification of bone marrow mesenchymal stem cells in diabetes treatment  Liu Jia 2015, 19 (28):  4549-4554.  doi: 10.3969/j.issn.2095-4344.2015.28.022 Abstract ( 252 )   PDF (1050KB) ( 238 )   Save BACKGROUND: Pancreas or islet cell transplantation and stem cell transplantation bring hope to cure diabetes, but pancreas or islet transplantation appears to have a lack of donors as well as immune rejection problems, limiting their clinical development. Therefore, stem cell transplantation therapy has become the current hotspot. OBJECTIVE: To study the therapeutic effects of huaman telomerase reverse transcriptase (hTERT)-modified bone marrow mesenchymal stem cells transplantation on diabetes mellitus in SD rats. METHODS: Bone marrow mesenchymal stem cells were transfected with PLXSN carrying hTERT. Thirty-six male SD rats were randomly divided into control group (n=6), stem cell group (n=10), hTERT transfection group (n=10), diabetes mellitus group (n=10). Except the control group, the rats were injected with stretozotocin (45 mg/kg) to make diabetes mellitus models. After modeling, rats in the stem cell group and hTERT transfection group were respectively intravenously injected with 1 mL of bone marrow mesenchymal stem cells (1.5×1010/L) and 1 mL of hTERT-modified bone marrow mesenchymal stem cells (1.5×1010/L). RESULTS AND CONCLUSION: At 24 hours after modeling, the fasting blood-glucose level was significantly increased in the diabetes mellitus group, which was higher than the normal value (6.7 mmol/L). At 15 days after cell transplantation, the fasting blood-glucose levels were signficiantly decreased in the stem cell group and hTERT transfection group as compared with the diabetes mellitus group (P < 0.05), but the body mass of rats was increased in these two group (P < 0.05), especially in the hTERT transfection group. At 45 days after cell transplantation, the fasting blood-glucose level and body mass in the stem cell group and hTERT transfection group were close to those in the control group (P > 0.05), and moreover, the hTERT group had better outcomes than the stem cell group. Meanwhile, in the diabetes mellitus group, the fasting blood-glucose level was still at a higher level, and the body mass decreased continously. These findings suggest that hTERT-modified bone marrow mesenchymal stem cell transplantation is effective for treatment of diabetes mellitus 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 Figures and Tables | References | Related Articles | Metrics

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Establishment and neural differentiation of spinocerebellar ataxia type 3-induced pluripotent stem cell lines Luo Min, Hu Dan, Niu Xiao-hua, Song Bing, Ou Zhan-hui, Fan Di, Wang Ding, He Wen-yin, Sun Xiao-fang 2015, 19 (28):  4555-4561.  doi: 10.3969/j.issn.2095-4344.2015.28.023 Abstract ( 448 )   PDF (6963KB) ( 335 )   Save BACKGROUND: Spinocerebellar ataxia type 3 (SCA3) is a typical genetic neurodegenerative disease. To establish patient-specific disease models of genetic background contributes to studying the pathogenesis and exploring therapeutic manners. OBJECTIVE: To observe the effectiveness of neural differentiation of induced pluripotent stem cell lines induced by SCA3 and the stability of CAG copy number. METHODS: Skin tissue of SCA3 patient was obtained clinically, and specific skin flbroblasts were isolated and   cultured. Reprogramming fibroblasts could obtain induced pluripotent stem cells. Patient-specific induced pluripotent stem cells, normal person induced pluripotent stem cells (NHF) and embryonic stem cells (ES-10) were induced to differentiate. Flow cytometry was used to compare the efficiency of differentiation. Western blot assay was utilized to detect ataxin-3 protein expression in neurons. Polymerase chain reaction was applied to measure the CAG repeat number of SCA3/ATXN3 gene. RESULTS AND CONCLUSION: Induced pluripotent stem cells that had identical genetic background to fibroblasts were successfully obtained, and had similar morphology and multi-directional differentiation potential to human embryonic stem cells. Each cell line could differentiate into neural stem cells. The CAG number did not apparently alter before and after reprogramming as well as induction of neuronal differentiation. The effectiveness of the differentiation of induced pluripotent stem cells derived from SCA3 into neural stem cells was lower than that of normal person-derived induced pluripotent stem cells (NHF) and embryonic stem cells (ES-10). These findings demonstrate that reprogramming can successfully establish human induced pluripotent stem cells, and induced the differentiation of above cells into neural stem cells. In the whole process, CAG number did not obviously alter, which was consistent with body cells of patients.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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In vivo homing of bone marrow mesenchymal stem cells transplanted through different ways in rats exposed to silica dust Huang Ming, Zhou Yong-mei, Yan Ling, Li Bin, Wu Qi-feng, Liang Wei-hui 2015, 19 (28):  4562-4566.  doi: 10.3969/j.issn.2095-4344.2015.28.024 Abstract ( 211 )   PDF (4764KB) ( 262 )   Save BACKGROUND: Bone marrow mesenchymal stem cells transplanted into rats exposed to silica dust can home to the injured lung, but the homing effects via different ways are still unclear. OBJECTIVE: To comparatively observe the distribution of bone marrow mesenchymal stem cells transplanted via different ways into rats exposed to silica dust.  METHODS: Bone marrow mesenchymal stem cells of donor rats were isolated through whole bone marrow adherent method and transfected by Lv-eGFP. Receptor rats were exposed to silica dust through windpipe injection and randomly divided into intravenous injection and intratracheal injection groups. Then, transfected bone marrow mesenchymal stem cells were injected via the vein and trachea into acceptor rats. The acceptor rats were killed at weeks 1, 2, 3, 4 after transplantation to take the lung, heart, liver, spleen, kidney, and brain tissue that were made into frozen sections and observed under fluorescence microscopy. The intensity of green  fluorescence (absorbance value) was analyzed using image analysis software. RESULTS AND CONCLUSION: Strong, wide and lasting green fluorescence was both observed in the lung tissue of intravenous injection and intratracheal injection groups, which was especially remarkable around the bronchus and blood vessels. The fluorescence intensities of both two groups were slightly decreased with time, but there was no difference between the two groups (P > 0.05). The fluorescence in the other organs of both two groups was also observed at early stage. It was stronger and wider in the liver, spleen and heart, while fainter and less in the kidney and brain, and reduced with time in all the organs. Fluorescence could be observed few and faint only in the liver and spleen at late stage, and could hardly be seen in the brain. The fluorescence intensities of the liver, spleen, heart, kidney and brain had no significant difference between the two groups at the same time (P > 0.05), but the fluorescence intensity in the brain at the 1st week showed significant difference between the two groups (P < 0.05). These findings indicate that the intravenous injection and intratracheal injection of bone marrow mesenchymal stem cells have similar homing effects in rats exposed to silica dust. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Umbilical cord blood mesenchymal stem cell transplantation for Parkinson’s disease: a feasibility study Liu Lei, Feng De-peng, Chen Yan, Zhao Xiu-min, Feng Xiao-ya, Ge Ru-cun, Xun Ying, Lv Yong-tao 2015, 19 (28):  4567-4571.  doi: 10.3969/j.issn.2095-4344.2015.28.025 Abstract ( 293 )   PDF (1457KB) ( 346 )   Save  BACKGROUND: Stem cells can be induced to differentiate into dopaminergic neurons in vivo and in vitro, which provides a theoretical basis for stem cell transplantation in the treatment of Parkinson’s disease OBJECTIVE: To explore the feasibility and mechanism of intracerebral transplantation of umbilical cord blood mesenchymal stem cells for treatment of Parkinson’s disease rats. METHODS: Intracerebral injection of 6-hydroxydopamine was used to make Parkinson’s disease models in SD rats. Twenty-two model rats were randomized into cell transplantation group (n=12) and control group (n=10) and respectively injected intracerebrally with umbilical cord blood mesenchymal stem cell suspension and PBS. At 1-8 weeks after cell transplantation, intra-abdominal injection of apomorphine was performed every week to observe the rotation behaviors of rats; at the 2nd and 8th weeks, rat’s striatum and substantia nigra were taken for immunohistochemistry staining. RESULTS AND CONCLUSION: The rotation behaviors were gradually decreased with time in the cell transplantation, but had no changes in the control group. At 3-8 weeks after transplantation, there were significant differences in the rotation behaviors between the two groups (P < 0.05). At 2 weeks after transplantation, tyrosine hydroxylase-positive cells were found within and around the striatum of the cell transplantation group; but there were no exogenous cells in the control group. At 8 weeks after transplantation,  there were still active cells and tyrosine hydroxylase-positive cells in the striatum of cell transplantation group, and there was no tyrosine hydroxylase expression in the striatum of the control group. These findings suggest that transplanted umbilical cord blood mesenchymal stem cells can survive in the brain that are positive for tyrosine hydroxylase, which can improve the behavior abnormalities of Parkinson’s disease 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 Figures and Tables | References | Related Articles | Metrics

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Autologous umbilical cord blood mononuclear cell transfusion in preterm children: immune function and prognosis Yang Chun-yan, Xu Ping, Li Bao-yun, Yang Yu-jun, Jia Huan-rong, Zhou Li-ying, Yang Qiao-zhi 2015, 19 (28):  4572-4575.  doi: 10.3969/j.issn.2095-4344.2015.28.026 Abstract ( 272 )   PDF (753KB) ( 274 )   Save BACKGROUND: The umbilical cord blood is rich in hematopoietic stem/progenitor cells that have strong proliferation and differentiation ability as well as ability to form colonies, and exert important roles in stimulating bone marrow function, improving blood cell viability and quantity, promoting immune cell maturation, and maintaining immune balance. OBJECTIVE: To evaluate the clinical effects of autologous umbilical cord blood mononuclear cell transplantation on the immunologic function and prognosis for premature infants. METHODS: Sixty-two preterm infants who entered into NICU immediately after birth, weighing ≤ 1 500 g, were divided into treatment group and control group according to parent’s willingness. In the treatment group, the umbilical cord blood was extracted from the umbilical vein and re-infused into the preterm infants after density gradient centrifugation within 4 hours. The cellular immunity levels, humoral immunity levels and clinical parameters were monitored before and after treatment. RESULTS AND CONCLUSION: After 1 week of treatment, the CD4, CD4/CD8 levels were significantly increased compared with the control group (P=0.01, 0.03), but CD8 level had no changes. At 1 week after treatment, IgM levels were both increased in the two groups, especially in the control group (P=0.00); IgA levels had no changes; IgG levels were decreased, especially in the control group (P=0.02). The incidence of severe infection during hospitalization was 13% in the treatment group, which was lower than the control group (16%), but there was no difference between the two groups. The proportion of infants undergoing mechanical ventilation and average length of stay had significant differences between the two groups (P < 0.05). After 12 months, the incidence of recurrent respiratory tract infections was zero in the treatment group and one case in the control group, and there was a significant difference between the two groups. These findings indicate that autologous umbilical cord blood mononuclear cell transplantation can improve the immunologic function, slower the reduction of IgG levels, reduce the usage of breathing machine, shorten the length of stay, and reduce the incidence of recurrent respiratory tract infections in preterm infants. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Influence of NGF-PEG-PLGA-NPs on neuronal differentiation of neural stem cells and PI3K/Akt signaling pathway    Chen Yan, Bao Guo-qing, Liu Fei-fei, Zhang Jun-du, Pan Cui-huan, Long Da-hong 2015, 19 (28):  4576-4581.  doi: 10.3969/j.issn.2095-4344.2015.28.027 Abstract ( 307 )   PDF (4632KB) ( 316 )   Save BACKGROUND: Our previous studies confirmed that NGF-PEG-PLGA-NPs has good sustained release effect and biological activity in vitro, and can induce the differentiation of PC12 cells into neuron-like cells. OBJECTIVE: To investigate the feasibility of neuronal differentiation of neural stem cells from septal area of fetal brain induced by NGF-PEG-PLGA-NPs and its influence on PI3K/Akt signaling pathway. METHODS: According to optimization prescription, NGF-PEG-PLGA-NPs were prepared by multiple emulsion solvent diffusion method. Neural stem cells were induced to neuronal differentiation in six groups, including control group, NGF group, NGF-PEG-PLGA-NPs group, LY294002 group, LY294002+NGF group and LY294002+NGF-PEG-PLGA-NPs group. Neurons were identified by immunofluorescence, while phosphorylation levels of Akt in PI3K/Akt signaling pathway were detected by western blotting. RESULTS AND CONCLUSION: The proportions of β-Tubulin III-positive neurons in control group, NGF group, NGF-PEG-PLGA-NPs group, LY294002 group, LY294002+NGF group and LY294002+NGF-PEG-PLGA-NPs group were (22.80±2.58)%, (35.80±3.98)%, (35.40±5.77)%, (26.60±3.87)%, (21.20±2.59)% and (25.80±7.22)%, respectively. There were no statistical differences in neuronal differentiation between NGF group and NGF-PEG-PLGA-NPs group (P > 0.05), but the ratios of neural differentiation in the two groups were both higher than that in the other four groups (P < 0.05). Western blotting results revealed that there were no statistical differences in Akt phosphorylation levels between NGF group and NGF-PEG-PLGA-NPs group (P > 0.05), but the phosphorylation levels of Akt were both higher than other four groups (P < 0.05). There were also no significant differences between LY294002+NGF and LY294002+NGF-PEG-PLGA-NPs groups and control group (P > 0.05), but the phosphorylation levels of Akt were higher than LY294002 group (P < 0.05). Results suggest that NGF-PEG-PLGA-NPs promoted neural differentiation of neural stem cells. The role might be related to increasing phosphorylation levels of Akt in PI3K/Akt signaling pathway. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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An air-liquid interface model of human lung epithelium generated from bronchiolar epithelial cells proliferated using medium containing ROCK kinase inhibitor  Jia Yuan-yuan, He Jin-xi, Sun Ying-fei, Han Fei, Yang Jia-li, Li Yong, Liu Xiao-ming 2015, 19 (28):  4582-4587.  doi: 10.3969/j.issn.2095-4344.2015.28.028 Abstract ( 321 )   PDF (5106KB) ( 246 )   Save BACKGROUND: Primary human lung epithelial cells are difficult to be isolated and cultured in vitro, which is characterized as limited sources, low cell viability, slow proliferation capacity, and lacking of differentiation capability. OBJECTIVE: To establish an air-liquid interface model of lung epithelium by in vitro proliferation of human bronchiolar epithelial cells, which is used for research on function of lung epithelial cells. METHODS: Primary human bronchiolar epithelial cells were isolated using Pronase and DNase I combined digestive methods, and then proliferated using medium containing ROCK kinase inhibitor. The proliferated cells were used for establishment of the air-liquid interface epithelium model. Cell differentiation was identified using scanning electron microscope, phase contrast microscope and immunofluorescent staining. RESULTS AND CONCLUSION: The primary human bronchiolar epithelial cells could be expanded successfully using medium containing ROCK kinase inhibitor, and the basal cell marker Cytokeratin14 was preferentially expressed in the proliferated cell population, indicating that these basal cells might be the main subpopulation of human lung epithelial stem cells. Subsequently, the proliferated cells under the air-liquid interface could differentiate into ciliated cells and non-ciliated column cells. The results suggest that the proliferation and differentiation of human bronchiolar epithelial cells were maintained in the presence of ROCK kinase inhibitor,  and the air-liquid interface could promote the differentiation of human bronchiolar epithelial cells.  BACKGROUND: Primary human lung epithelial cells are difficult to be isolated and cultured in vitro, which is characterized as limited sources, low cell viability, slow proliferation capacity, and lacking of differentiation capability. OBJECTIVE: To establish an air-liquid interface model of lung epithelium by in vitro proliferation of human bronchiolar epithelial cells, which is used for research on function of lung epithelial cells. METHODS: Primary human bronchiolar epithelial cells were isolated using Pronase and DNase I combined digestive methods, and then proliferated using medium containing ROCK kinase inhibitor. The proliferated cells were used for establishment of the air-liquid interface epithelium model. Cell differentiation was identified using scanning electron microscope, phase contrast microscope and immunofluorescent staining. RESULTS AND CONCLUSION: The primary human bronchiolar epithelial cells could be expanded successfully using medium containing ROCK kinase inhibitor, and the basal cell marker Cytokeratin14 was preferentially expressed in the proliferated cell population, indicating that these basal cells might be the main subpopulation of human lung epithelial stem cells. Subsequently, the proliferated cells under the air-liquid interface could differentiate into ciliated cells and non-ciliated column cells. The results suggest that the proliferation and differentiation of human bronchiolar epithelial cells were maintained in the presence of ROCK kinase inhibitor,  and the air-liquid interface could promote the differentiation of human bronchiolar epithelial cells.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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MRI tracing of stem cells: theory and application progress Long Teng-he, Cui Hui-qin 2015, 19 (28):  4588-4592.  doi: 10.3969/j.issn.2095-4344.2015.28.029 Abstract ( 347 )   PDF (860KB) ( 247 )   Save BACKGROUND: Continuous monitoring is required in the survival, distribution, migration, proliferation and differentiation of transplanted stem cells in vivo thereby to assess its efficacy and safety. OBJECTIVE: To analyze the characteristics of literature addressing stem cell markers and MRI tracer in the past 10 years. METHODS: The first author retrieved CNKI and Web of Science by computer for articles about MRI tracing of stem cells published from January 2005 to December 2014, using the keywords of “stem cells, magnetic resonance imaging, tracing technique” that appeared in the title and abstract. Then, the retrieved articles were further analyzed. RESULTS AND CONCLUSION: Currently, tracing technique has been widely used in stem cell therapy, organ transplantation, bacterial infection, gene expression and new drug development. Stem cell tracing technologies include isotope tracing, antigenic labeling, fluorescence protein labeling, fluorescent dye labeling, nuclear magnetic resonance contrast enhancer labeling, Lac-Z gene labeling, Y chromosome labeling. Recently, with the development of molecular imaging, MRI tracing techniques have been increasingly used to assess the efficacy and safety of stem cell 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 Figures and Tables | References | Related Articles | Metrics