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28 March 2018, Volume 22 Issue 9 Previous Issue    Next Issue

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Bone defect repair using allogenic bone carrying bone morphogenetic protein-2 and gene transfected bone marrow mesenchymal stem cells Wang Xiao-zhi, Yang Hua, Zhang Chen, He Hui-yu, Ba Jiao-jiao 2018, 22 (9):  1313-1318.  doi: 10.3969/j.issn.2095-4344.0458 Abstract ( 359 )   PDF (2465KB) ( 284 )   Save BACKGROUND: Bone tissue engineering technology brings a new path to treat bone defects, and moreover, gene transfection optimizes the effects of tissue-engineered bone in bone repair. OBJECTIVE: To observe the effect of allogenic bone carrying bone morphogenetic protein-2 (BMP-2) and basic fibroblast growth factor (bFGF) transfected bone marrow mesenchymal stem cells (BMSCs) in the repair of critical-sized bone defects of the sheep ilium. METHODS: Nine Altay sheep were taken to make animal models of critical-sized bone defects in the bilateral ilia. Allogenic bone carrying BMP-2 and bFGF-transfected BMCSCs was implanted into the left side of the ilium (n=9, experimental group), while allogenic bone carrying BMSCs (n=3, cell scaffold group), allogenic bone (n=3, allogenic bone group) and β-tricalcium phosphate (n=3, β-TCP group) were implanted into the right side of the ilium, respectively. Histological observation and immunohistochemical staining were performed at 4, 8, 12 weeks after implantation. RESULTS AND CONCLUSION: Findings from histological observation showed that: at 12 weeks after implantation, the experimental group exhibited obvious bone remodeling, largely absorbed bone allograft, gradually reduced osteoclast-like cells and increased osteoblast-like cells, clear trabecular structure, dense bone matrix around the trabecular bone, and large area of flaky new bone tissues. Bone materials also degraded in the other three groups, but no remodeling occurred. Immunohistochemical staining showed that at 12 weeks after implantation, bone sialoprotein type I collagen in the experimental group were strongly positive, while they were weakly expressed in the remaining three groups. Overall, our findings indicate that the allogenic bone carrying BMP-2 and bFGF-transfected BMSCs can thoroughly repair critical-sized bone defects of the sheep ilium.  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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A comparative study on the secretion of various cytokines by human adipose-derived stem cells at different passages Wang Qian, Tang Wen-yan, Wang Zhao-yan, Yang Yin-xiang, Luan Zuo 2018, 22 (9):  1319-1324.  doi: 10.3969/j.issn.2095-4344.0459 Abstract ( 575 )   PDF (1522KB) ( 272 )   Save BACKGROUND: Adipose-derived stem cells (ADSCs) can establish a favorable repair microenvironment by secreting abundant cytokines, which allows ADSCs to be a good source of seed cells for the treatment of ischemic diseases. OBJECTIVE: To investigate the changes of cytokines secreted by human ADSCs at passages 2-10. METHODS: After isolation and culture of ADSCs from human adipose tissue, the morphological features of cells were observed under inverted microscope. Human ADSCs were identified by the immunophenotypes and differentiation capability. RESULTS AND CONCLUSION: ADSCs were fusiform or polygonal in shape, with buging cell body, homogenized cytoplasm and clear nuclei, and could differentiate into adipocytes, osteocytes and chondroblasts in vitro. ADSCs at passage 3 were positive for CD29 (99.21%), CD73 (99.65%) and CD90 (99.92%), but negative for hematopoietic marker CD34 (2.25%). ELISA results showed that ADSCs at passage 5 had the highest secretion levels of vascular endothelial growth factor and hepatocyte growth factor, while ADSCs at passage 3 had the highest secretion level of brain-derived neurotrophic factor. To conclude, ADSCs have steady biological features of stem cells and exhibit good growth and proliferation potentials. ADSCs at different passages have different capacities in the secretion of vascular endothelial growth factor, hepatocyte growth factor and brain-derived neurotrophic factor. Passage 5 ADSCs show the highest ability to secrete vascular endothelial growth factor and hepatocyte growth factor, while passage 3 ADSCs show the strongest potential to secrete brain-derived neurotrophic factor. 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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Ascorbic acid influences on extracellular matrix and structure of rabbit bone marrow mesenchymal stem cells Yao Zhi-ye, Liu Yu-mei, Chen Yan-ling, Chen Liang, He Shao-ru, Zhang Zhan-song 2018, 22 (9):  1325-1331.  doi: 10.3969/j.issn.2095-4344.0460 Abstract ( 392 )   PDF (5206KB) ( 294 )   Save BACKGROUND: The effect of extracellular matrix on stem cells is the focus of tissue engineering. However, there are few reports about the synthesis and secretion of extracellular matrix as well as its effects on cells. OBJECTIVE: To isolate, culture and identify rabbit bone marrow mesenchymal stem cells (BMSCs), and to explore the changes of extracellular matrix and whole structure under the intervention of ascorbic acid. METHODS: Rabbit BMSCs were isolated by differential adherent method of the bone marrow, and the expression of CD44, CD45 and CD31 was identified by flow cytometry. The BMSCs were cultured in the culture medium containing 20 mg/L ascorbic acid. Then the cell morphology, gross structure, ultrastructure, and histological changes of BMSCs were observed. The expression of extracellular matrix related genes was detected by RT-PCR. RESULTS AND CONCLUSION: Over 95% passage 2 BMSCs could express CD44, but the expression levels of CD45 and CD31 were extremely low. Intervention with ascorbic acid enhanced the proliferation of BMMSCs with unclear cell boundaries. A cell-sheet structure formed at 10-14 days after intervention. Hematoxylin-eosin staining results showed a layered cell arrangement, and Masson staining findings showed a large amount of extracellular matrix composition. Abundant endoplasmic reticula and vesicle-like structure were observed under the transmission electron microscope. RT-PCR findings showed that ascorbic acid significantly increased the expression of fibronectin mRNA in the BMSCs (P < 0.05), but slightly increased the mRNA expression of collagen type I. All these findings indicate that ascorbic acid not only increases the proliferation and transformation of rabbit BMSCs, but also promotes the synthesis and secretion of extracellular matrix, which has great potential in tissue engineering applications. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Diazoxide effects on proliferation and apoptosis of bone marrow mesenchymal stem cells under hypoxic conditions Li Hong-bo, Lin Shi-wei, Yao Jiang-ling, Bian Yang-yang, Qu Ye, Wang Lei-tao, Peng Lei 2018, 22 (9):  1332-1337.  doi: 10.3969/j.issn.2095-4344.0461 Abstract ( 301 )   PDF (5408KB) ( 256 )   Save BACKGROUND: It is an urgent problem to effectively make bone marrow mesenchymal stem cells exert proper effects under hypoxic preconditioning. OBJECTIVE: To investigate the effects of diazoxide, a Mito-KATP channel activator, on the proliferation and apoptosis of mouse BMSCs in hypoxic environment. METHODS: Mouse BMSCs were divided into four groups: blank control group, 0.16, 0.8, 4 μmol/L diazoxide groups. Cells intervened by diazoxide were cultured in a 10% O2 incubator. MTT assay was performed to detect cell proliferation at 1, 2, 4, 6, 8 days after intervention, and Hoechst 33258 staining was performed to observe cell apoptosis at 14 days after intervention.  RESULTS AND CONCLUSION: High homogeneity and purity but low proliferation of BMSCs was found. There was no significant difference in the activity of BMSCs among 0.16, 0.8, 4 μmol/L diazoxide groups (P > 0.05). In the blank control group, concentrated nuclei were dark blue in color and aggregated, and several round apoptotic bodies were found. In the diazoxide groups, apoptotic bodies were occasionally found, and no significant difference was found among different diazoxide groups. These findings indicate that a certain concentration of diazoxide can reduce cell apoptosis but has no effects on the proliferation of mouse BMSCs under hypoxic environment (10% O2). 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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Repair of large-segmental femoral defects using lentivirus-mediated bone morphogenetic protein 2/bone marrow mesenchymal stem cells/demineralized bone matrix Tao Xuan, Li Qiang, Li Shi-peng, Shi Zheng-song, Zhou Zhen-jie 2018, 22 (9):  1338-1343.  doi: 10.3969/j.issn.2095-4344.0462 Abstract ( 307 )   PDF (4822KB) ( 299 )   Save BACKGROUND: Our research team has confirmed that compared to the adenoviral vector, transfection by lentiviral vector to rabbit bone marrow mesenchymal stem cells (BMSCs) is more effective that the expression of enhanced green fluorescent protein (EGFP)/bone morphogenetic protein 2 (BMP-2) can be persistent for a longer term. But further investigations are needed to explore whether BMSCs transfected with hBMP-2 through lentivirus combined with demineralized bone matrix (DBM) can promote bone defect repair. OBJECTIVE: To evaluate the effect of lentivirus-mediated hBMP-2/BMSCs/DBM (LV-hBMP-2/BMSCs/DBM) on the repair of large-segmental femoral defects and to explore the new treatments for large-segmental femoral defects. METHODS: Large-segmental bone defect models were made in the right femur of 48 New Zealand white rabbits by cutting the middle femoral bone and steel plate fixation. Then, animal models were randomly divided into four groups (n=12 per group) and implanted with nothing (control), DBM, hBMP-2/DBM, and LV-hBMP-2/BMSCs/DBM. Three rabbits from each group were sacrificed at 2, 4, 8 and 12 weeks after surgery to evaluate the repairing effect of femoral defects through hematoxylin-eosin staining, biomechanical analysis and radiological examination. RESULTS AND CONCLUSION: X-ray results revealed that osteotylus formed in all the four groups to different extents, and Lane - Sandhu X-ray scores were ranked as follows: control group < DBM group < hBMP-2/DBM group < LV-hBMP-2/BMSCs/DBM group (P < 0.05). Findings from the three-point bending test showed that the maximum load of the LV-hBMP-2/BMSCs/DBM group was significantly higher than that of the hBMP-2/DBM group, but is still lower than that of the normal femur at 8 and 12 weeks after modeling (P < 0.05). Hematoxylin-eosin staining results revealed that a few trabecular bones arranged disorderedly and a large amount of fibrous tissues in the control group; the DBM scaffold was basically degraded in the DBM group presenting with partially disordered trabecular bones and a large amount of fibrous tissues; the trabecular bones in the bone defect area were basically connected into line to start the shaping of the cortical bone, and recanalization of the medullary cavity was insignificant in the hBMP-2/DBM group; new cortical bone formed in the bone defect area and the medullary cavity was recanalized in the LV-hBMP-2/BMSCs/DBM group. These findings indicate that LV-hBMP-2/BMSCs/DBM can produce a large amount of calluses, promote formation of new cortical bone, and promote bone conduction, bone induction and osteogenesis after implantation into the bone defect; and this material has good repairing effect on large-segmental femoral defects of rabbits. 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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Cardioprotection by hypoxia-induced rat adipose-derived stem cells through paracrine mechanisms Gao Yu-ping, Lin Yuan-yuan, Li Xue-wen, Fan Chun-hui, Yang Fan, Hao Da-jie, Ge Wen-jia 2018, 22 (9):  1344-1349.  doi: 10.3969/j.issn.2095-4344.0752 Abstract ( 430 )   PDF (876KB) ( 215 )   Save BACKGROUND: Adipose-derived stem cells (ADSCs) represent one of the promising cell sources for myocardial regeneration due to their easy accessibility and efficacy in the improvement of cardiac function following myocardial infarction. However, previously reported studies on the underlying mechanism of ADSCs-mediated cardioprotective effect mainly focused on the ADSCs cultured at room air. OBJECTIVE: To test the paracrine actions and anti-apoptotic effect of ADSCs under hypoxic conditions. METHODS: After isolation and culture, neonatal rat myocardial cells were injured by hydrogen peroxide and co-cultured with rat ADSCs under normoxia and hypoxia (10% O2) conditions. Ratio of reduced glutathione to oxidized glutathione (GSH/GSSG) in the cell pellet and levels of vascular endothelial growth factor (VEGF), insulin-like growth factor 1 (IGF-1), and basic fibroblast growth factor (bFGF) were tested by ELISA. Expression of apoptotic proteins Bax and Bcl-2 were determined by western blot. RESULTS AND CONCLUSION: GSH/GSSG, VEGF, IGF-1, and bFGF were decreased in neonatal rat myocardial cells injured by hydrogen peroxide. ADSCs significantly attenuated hydrogen peroxide-induced myocardial apoptosis by increasing the ratio of GSH/GSSG and the secretion of VEGF, IGF-1 and bFGF. ADSCs also down-regulated Bax expression and up-regulated Bcl-2 expression. To conclude, hypoxic conditions can enhance the anti-apoptosis and cardioprotective effects of ADSCs through the paracrine mechanism. 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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Risk factors for hepatocellular carcinoma in hepatitis B cirrhosis patients given treatment with human umbilical cord mesenchymal stem cell Ren Hao, Dong Jing, Liu Li-wei, Chen Zhao-lin, Pan Jin-jin, Chen Xi, Song Hai-yan, Zhang Jun-fei, Chen Cong-xin, Liu Bo 2018, 22 (9):  1350-1356.  doi: 10.3969/j.issn.2095-4344.0463 Abstract ( 332 )   PDF (1495KB) ( 273 )   Save BACKGROUND: With the continuous development of mesenchymal stem cell therapy, it has been reported that stem cell therapy is likely to cause the occurrence and development of tumors. OBJECTIVE: To investigate the potential risks of hepatocellular carcinoma (HCC) in patients with hepatitis B cirrhosis after receiving human umbilical cord mesenchymal stem cells (hUC-MSCs) transplantation. METHODS: The study collected the information of patients with hepatitis B cirrhosis treated with hUC-MSCs, admitted at the Infectious Disease Department of the 105th Hospital of PLA from January 2011 to December 2013. The following investigation lasted 36 months. The follow-up was terminated at the time of diagnostic confirmation. The risk factors that may affect the occurrence of HCC were analyzed by univariate Logistic and multivariate unconditional Logistic regression analyses. RESULTS AND CONCLUSION: (1) A total of 386 patients were followed up, including 171 patients who received hUC-MSCs transplantation as the observation group and 215 patients only given general internal medicine treatment as the control group. (2) At the follow-up of 12 months, the incidence of HCC in the observation group was significantly higher than that in the control group (P < 0.05). At the follow-up of 36 months, the incidence of HCC was 11.7% in the observation group and 9.8% in the control group (P > 0.05). (3) Univariate Logistic regression analysis showed that the HCC patients had higher age, alpha-fetoprotein (AFP), alpha-fetoprotein variants (AFP-L3), AFP-L3 ratio (AFP-L3%), and Golgi glycoprotein 73 (GP73) than those with no HCC in both control and observation groups (P < 0.05). Multivariate unconditional Logistic regression analysis showed that only APF-L3% was an independent risk factor for HCC in patients with hepatitis B cirrhosis undergoing hUC-MSCs transplantation. Overall, hUC-MSCs transplantation does not increase the HCC incidence in patients with hepatitis B cirrhosis within 3 years, but it may lead to an early onset of HCC. AFP-L3% can be used as an early predictor of HCC in patients with hepatitis B cirrhosis undergoing hUC-MSCs 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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Induced differentiation of adipose-derived mesenchymal stem cells into Leydig cells Fu Jin-shan, Liang Pei-yu, Ou Shan-ji 2018, 22 (9):  1364-1369.  doi: 10.3969/j.issn.2095-4344.0465 Abstract ( 483 )   PDF (4351KB) ( 268 )   Save BACKGROUND: The recent development of stem cells has provided new ideas for the treatment of androgen-deficient diseases. OBJECTIVE: To investigate whether adipose-derived mesenchymal stem cells can differentiate into Leydig cells. METHODS: Passage 3 rat adipose-derived mesenchymal stem cells that grew well were taken and cultured in the medium with (experimental) or without (control) 0.1 mg/L human chorionic gonadotropin, 10.0 μg/L platelet-derived growth factor and 10.0 μg/L basic fibroblast growth factor. Indicator detection was done at 1, 7, 14, 24 days of induced culture. RESULTS AND CONCLUSION: (1) Immunofluorescence staining results showed that there were no 3β-hydroxysteroid dehydrogenase (3β-HSD) positive cells in the control group, while the number of 3β-HSD positive cells was gradually increased in the experimental group with the induction time, which presented with fluorescence enhancement. (2) There was no secretion of testosterone in the control group, while in the experimental group, testosterone secretion was detected at 7 days of induced culture, and moreover, the testosterone level was increased with the induction time. (3) RT-PCR findings showed no luteinizing hormone receptor, steroidogenic acute regulatory protein, and 3β-HSD positive bands in the control group, while these positive bands appeared in the experimental group after 1 day of induction, and strengthened with the induction time. To conclude, adipose-derived mesenchymal stem cells can be induced to differentiate into Leydig 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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Effect of 1,25(OH)2D3 on type I collagen secretion in adipose-derived mesenchymal stem cells and its mechanism Zhai Min, Hu Xiao-gen, Liu Hong-lin, Xu Shi-qing, Wang Zai, Zhang Wen-jian 2018, 22 (9):  1370-1375.  doi: 10.3969/j.issn.2095-4344.0466 Abstract ( 345 )   PDF (5354KB) ( 241 )   Save BACKGROUND: Adipose-derived mesenchymal stem cells (ADMSCs) have been reported to improve wound healing. However, type I collagen secreted by ADMSCs will contribute to scar formation. Therefore, inhibiting type I collagen secretion from ADMSCs will strengthen its clinical application. OBJECTIVE: To investigate the effect of 1,25(OH)2D3 on secretion of type I collagen by ADMSCs and its mechanism. METHODS: Human ADMSCs were isolated by collagenase digestion, and identified by flow cytometry. ADMSCs at passage 4 were cultured in DMEM/F12 medium containing different concentrations of 1,25(OH)2D3 (10-7, 10-8, 10-9, 10-10 and 0 mol/L) respectively for 4 days. Then, the concentration of type I collagen in cell supernatant was measured by ELISA. Real-time PCR and western blot were used to detect the expression of Smad3 at mRNA and protein levels and phosphorylated protein Smad3 level in ADMSCs cultured with and without 1,25(OH)2D3. To analyze the contribution of Smad3 to the effect of 1,25(OH)2D3, Smad3 inhibitor was added to culture medium 30 minutes before adding 1,25(OH)2D3, and type I collagen in cell supernatant was detected by ELISA at 4 days after addition of SMAD3 inhibitor. RESULTS AND CONCLUSION: 1,25(OH)2D3 inhibited the secretion of type I collagen by ADMSCs in a dose-dependent manner. The results of real-time PCR and western blot showed that the expression of Smad3 was upregulated by 1,25(OH)2D3, and the results of western blot showed that the phosphorylated Smad3 protein level in ADMSCs was significantly increased by 1,25(OH)2D3. Moreover, the inhibition of type I collagen secretion by 1,25(OH)2D3 could be blocked by Smad3 inhibitor. These results indicate that 1,25(OH)2D3 can inhibit the secretion of type I collagen from ADMSCs by up-regulating the expression of Smad3. 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 leukemia inhibitory factor receptor overexpression on stemness maintenance and lung metastasis in vivo of thyroid cancer stem cells Zhang Zhen-hua, Su Zi-jie, Kan Yun-zhen, Liu Qiu-yu 2018, 22 (9):  1376-1381.  doi: 10.3969/j.issn.2095-4344.0467 Abstract ( 381 )   PDF (4759KB) ( 244 )   Save BACKGROUND: Thyroid cancer stem cells are essential to the recurrence and metastasis of thyroid carcinoma. Leukemia inhibitory factor receptor (LIFR) shows a downward trend in a variety of malignant tumors, and its overexpression can inhibit the recurrence and metastasis of malignant tumors. OBJECTIVE: To explore the effect of LIFR on the stemness maintenance and lung metastasis of thyroid cancer stem cells in vivo.  METHODS: Primary thyroid cancer cells TCLM were isolated from the lung metastases of a metastatic thyroid cancer patient. Serum-free suspension culture was used to form tumor cell balls. Flow cytometry was used to screen CD133+ phenotype of metastatic thyroid cancer stem cell subpopulation TCLM-S. The overexpressed recombinant lentiviral plasmid containing LIFR and its negative control containing the empty plasmid were infected into thyroid cancer stem cells TCLM-S at the ratio of virus/cell number=20, and screened with 2.0 mg/L puromycin to construct TCLM-SLIFR and TCLM-Scontrol stem cells which stably expressed LIFR and its control. Real-time quantitative PCR (qRT-PCR) was used to detect the expression of LIFR in TCLM-SLIFR and TCLM-Scontrol stem cells. Flow cytometry was used to detect the percentage of CD133+ phenotype cell subsets, western blot assay was used to detect the expression of tumor stemness related factors SOX2, Oct4, Nanog and tumor invasion and metastasis related proteins E-cadherin, matrix metalloproteinase (MMP)-2, MMP-7 in TCLM-SLIFR and TCLM-Scontrol stem cells. TCLM-SLIFR and TCLM-Scontrol stem cells were respectively injected into BALB/c nude mice by tail vein, and the lung metastasis model of thyroid cancer stem cells was constructed. The effect of LIFR overexpression on lung metastasis was observed. RESULTS AND CONCLUSION: Compared with TCLM-Scontrol cells, the expression of LIFR in TCLM-SLIFR cells was significantly increased, the proportion of CD133+ phenotype stem cell subsets was significantly decreased, the expression of SOX2, Oct4 and Nanog were significantly decreased, the expression of E-cadherin was significantly increased, and the expression of MMP-2 and MMP-7 was significantly decreased. Moreover, the number of lung metastasis in nude mice given TCLM-SLIFR cells was significantly decreased as compared with those given TCLM-Scontrol cells. To conclude, LIFR overexpression can decrease the stemness and ability of lung metastasis in vivo. 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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Comparison of exosome extracting methods from human umbilical cord mesenchymal stem cells Guo Ying, Wang Xiu-wei, Niu Yu-hu, Wang Li, Zhou Nan, Li Bai-yi, Wang Zhen-dong, Zhang Pin, Gao Ya-jie, Niu Bo 2018, 22 (9):  1382-1388.  doi: 10.3969/j.issn.2095-4344.0468 Abstract ( 1169 )   PDF (5231KB) ( 371 )   Save BACKGROUND: Cell-free stem cell therapy has been an issue of concern, but there is no conclusion on how to extract high-quality exosomes. OBJECTIVE: To extract exosomes from human umbilical cord mesenchymal stem cells by using three different methods, and then to screen the optimal method. METHODS: Exosomes were extracted from human umbilical cord mesenchymal stem cells by using the Total Exosome Isolation test kit, Exo Quick test kit and differential ultracentrifugation method, respectively. Then, transmission electron microscopy was used for morphological observations, BCA was utilized to quantify the protein, and western blot assay was applied to detect surface markers CD9, CD81 and CD63. RESULTS AND CONCLUSION: Extraction of exosomes was completed by all the three methods, and round or oval membranous vesicles were observed under the transmission electron microscope. The protein content and purity of exosomes was highest in the differential ultracentrifugation group, followed by the Exobiology Quick kit group, and lowest in the Total Exosome Isolation kit group, and there were significant differences among the three groups (P < 0.05). Under the same protein concentration, surface specific markers, CD81, CD63 and CD9, were expressed highest in the differential ultracentrifugation group, followed by the Exobiology Quick kit group, and lowest in the Total Exosome Isolation kit group. The operating time was significantly lower in the Exobiology Quick kit group compared with the other two groups (P < 0.05). To conclude, despite a longer operating time, the differential ultracentrifugation method is a rational method to extract enough exosomes with relative high purity.  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 bone marrow mesenchymal stem cell transplantation on voltage-gated   K+ channel proteins and cytokines in the infarcted myocardium of rats Hu Ji-hong, Zhao Jing-miao, Wang Qiu-ping, Jia Jia, Lu Juan, Jin Hua, Hou Qian 2018, 22 (9):  1389-1394.  doi: 10.3969/j.issn.2095-4344.0469 Abstract ( 343 )   PDF (7367KB) ( 196 )   Save BACKGROUND: Studies have shown that bone marrow mesenchymal stem cell (BMSC) transplantation can effectively improve cardiac function after myocardial infarction. However, few reports have been issued on myocardial electrophysiology after BMSCs transplantation.  OBJECTIVE: To observe the effects of BMSCs transplantation on voltage-gated K+ channel protein and myocardial infarction-related cytokines, thereby providing basic evidence for further exploration on the mechanism underlying arrhythmia in myocardial infarction due to BMSCs transplantation.  METHODS: Forty male Wistar rats, SPF grade, were randomly divided into four groups: sham group, model group, cell culture medium group and BMSCs group. The myocardial infarction model was created in rats by permanent ligation of the left descending coronary artery. At 15-20 minutes after surgery, BMSCs (100 µL, 1×106) or cell culture medium (100 µL) was injected at four sites in the peri-infarct zone. Four weeks after cell therapy, cardiac samples were taken, the pathological morphology of the infarcted myocardium was observed by hematoxylin-eosin staining, and the infarct size was calculated; the expression levels of voltage-gated K+ channel proteins Kv1.2 and Kv1.5 and cardiac troponin T (cTnT) were measured by western blot assay; and the expression levels of apoptotic factor (Caspase-3), autophagy factor (Bcl-2), nitric oxide and superoxide dismutase were tested by immunohistochemistry.  RESULTS AND CONCLUSION: Compared with the model group and cell culture medium group, the infarct size decreased in the BMSCs group (P < 0.05); the expression levels of cTnT, Kv1.5 and superoxide dismutase increased (P < 0.05), and the expression levels of Caspase-3, Bcl-2 and Kv1.2 decreased (P < 0.05) in the BMSCs group. In summary, BMSCs transplantation can promote the expression of voltage-gated K+ channel proteins, and improve anti-oxidation capacity of the myocardium and decrease apoptosis and autophagy.  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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Transplantation of umbilical cord blood mesenchymal stem cells at two states: comparison of neurological function recovery in cerebral infarction rats Han Xiao-gai, Li Jian-bin, Shan Hong, Bie Li-li, Wang Jiao-jie, Liu Ying, Qi Zheng 2018, 22 (9):  1395-1401.  doi: 10.3969/j.issn.2095-4344.0470 Abstract ( 394 )   PDF (3013KB) ( 224 )   Save BACKGROUND: Human umbilical cord blood mesenchymal stem cells (hUC-MSCs) can repair the injury of nerve cells caused by ischemia and hypoxia, but which state of cells has a better therapeutic efficacy, primary isolation or induced differentiation is not yet known. OBJECTIVE: To compare the therapeutic effects of hUC-MSCs primary cultured and differentiated in a rat model of cerebral infarction. METHODS: After full-term delivery, fetal umbilical cord blood samples were obtained using quadruple bags by means of density gradient centrifugation. hUC-MSCS were induced in the medium containing basic fibroblast growth factor. Sixty rats were equivalently randomized into four groups: sham, model, primary culture and induced differentiation groups. Animal models of cerebral infarction were made in the rats in the latter three groups. Model rats in the primary culture and induced differentiation groups were subjected to tail vein transplantation of hUC-MSCs that were primary cultured or induced to differentiate in vitro at 7 days after modeling.  RESULTS AND CONCLUSION: A marked improvement in balance, walking, spatial orientation, and learning and memory abilities was found in the rats after transplantation of hUC-MSCs that were primary cultured or induced to differentiate. Moreover, compared with the primary culture group, a significant improvement was found in the induced differentiation group, including improved pathological injury of the brain, higher expression of CD34 and Ki-67, lower expression of glial fibrillary acidic protein, lower expression of interleukin 1β and tumor necrosis factor β. Compared with the primary culture group, similar infarction size and expression of interleukin-6 were also found in the induced differentiation group. These findings indicate that hUC-MSCs with induced differentiation exhibit better therapeutic outcomes in the recovery of neurological function of cerebral infarction 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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Feasibility of transplanting human bone marrow mesenchymal stem cells into manganese poisoning rats Guo Shu-han, Song Ping-ping, Chen Ceng-ceng, Tian Yu-tian, Fan Xiao-li, Yan Yong-jian 2018, 22 (9):  1402-1406.  doi: 10.3969/j.issn.2095-4344.0471 Abstract ( 304 )   PDF (3544KB) ( 185 )   Save BACKGROUND: Studies have shown that bone marrow mesenchymal stem cells (BMSCs) can treat central nervous system diseases. BMSCs have the function of self-renewal and differentiation into a variety of neural cell types. BMSCs with self-renewal and multi-directional differentiation abilities can successfully differentiate into dopaminergic neurons after transplantation into an animal model. OBJECTIVE: To observe the effect of transplanted BMSCs on behavior and dopaminergic neurons in rats with manganese poisoning. METHODS: Rat models of manganese poisoning were constructed by intraperitoneal injection of MnCl2•4H2O into Sprague-Dawley rats. The model rats were then randomly divided into two groups, BMSCs and phosphate buffered solution (PBS) control group, and 5 μL of passage 3 human BMSCs suspension or equivalent PBS was transplanted into the right striatum of the manganese poisoning rats. One month after transplantation, the rats were subjected to behavioral assessment. The differentiation of BMSCs was observed by immunofluorescence. The contents of dopamine, brain-derived neurotrophic factor, glial cell-derived neurotrophic factor in the right striatum of rats were detected by ELISA. RESULTS AND CONCLUSION: The behavioral score of the BMSCs treated group was significantly lower than that of the PBS control group after transplantation (P < 0.05). Double-labeled positive cells for human-specific nuclear antigen/tyrosine hydroxylase (hNUC/TH) and human-specific nuclear antigen/glial cell-derived acidic protein (hNUC/GFAP) were observed in the BMSCs treated group after transplantation. Meanwhile, hNUC/TH and hNUC/GFAP double-labeled positive cells were undetected in the PBS control group after transplantation. The expression levels of dopamine, brain-derived neurotrophic factor, glial cell-derived neurotrophic factor in the BMSCs treated group were higher than those in the PBS control group. This suggests that BMSCs can improve the behavior of manganese poisoning rats and can differentiate into dopaminergic neurons and astrocytes. 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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Human umbilical cord mesenchymal stem cells in the treatment of knee osteoarthritis: study protocol for a clinical trial Yang Zi-yi, Lin Jian-hao, Xing Dan, Wang Bin, Hou Yun-fei 2018, 22 (9):  1407-1412.  doi: 10.3969/j.issn.2095-4344.0472 Abstract ( 856 )   PDF (803KB) ( 386 )   Save BACKGROUND: Osteoarthritis (OA) is a kind of chronic bone and joint disease which seriously endangers human health. Cell therapy for OA has aroused widespread concern and gotten rapid development in recent years. Umbilical cord-derived mesenchymal stem cells (UC-MSCs) have the advantages of easy amplification and differentiation, anti-inflammation and recruiting function such as MSCs from other sources. Furthermore, UC-MSCs are young cells that have large quantity, no ethical problems, high proliferative potential and pluripotent differentiation. UC-MSCs have been the most commonly used seed cells in clinical cell therapy. OBJECTIVE: To evaluate the efficacy and safety of UC-MSCs in the treatment of human knee OA to provide theoretical and clinical basis for stem cell therapy of OA. METHODS: The trail will be completed in Arthritis Clinic & Research Center, Beijing, China. Participants will be recruited according to established inclusion/exclusion criteria after obtaining the informed consent and the approval of the Ethics Committee (the first and second parts of the trial have been registered (https://register.clinicaltrials.gov/), with the identifier No. NCT03357770 and NCT03358654, and the third part will be carried out according to the conclusion of the first and second parts). The clinical trial will be divided into three parts: in the first part three groups will be recruited. Each group will contain three participants. The three groups of participants will be treated with high, medium and low dose of MSCs, respectively. Participants will be followed up to evaluate dose-limiting toxicity so as to determine the maximum tolerated dose. The second part will be a single-arm clinical trial. Nine participants will be recruited. The injection dose will be the maximum tolerated dose determined in the first part. Participants will be followed up to evaluate the safety and efficacy of the treatment. The third part will be a randomized controlled trial. Participants will be randomly divided into two groups (n=7 per group) and treated with MSCs and hyaluronic acid, respectively. During the trial, evaluators, participants and interveners will be unaware of grouping information and interventions. Participants will be followed up at designed time points after treatment to evaluate the safety and efficacy of the intervention. The trial will be terminated if there are unexplained local and systemic symptoms or death according to the NCI-CTCAE criteria. EXPECTED RESULTS: With reference to the previous literature, the knee pain will be relieved, the score of knee joint function will increase, and the cartilage defect area will decrease on MRI at 1-2 years after the intervention. The trail is expected to spend 3 years and 6 months. 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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Peripheral blood haploidentical hematopoietic stem cell transplantation for treatment of acute lymphoblastic leukemia in adults: monitoring minimal residual diseases to intervene recurrence Wan Ding-ming, Sun Lin-lin, Xie Xin-sheng, Guo Rong, Cao Wei-jie, Zhang Su-ping, Li Li, Chen Xiao-na, Liu Yu-ye 2018, 22 (9):  1413-1418.  doi: 10.3969/j.issn.2095-4344.0473 Abstract ( 392 )   PDF (4334KB) ( 206 )   Save BACKGROUND: Recurrence of acute leukemia after hematopoietic stem cell transplantation is one of the major problems affecting the long-term survival of patients. Early intervention to prevent ALL recurrence after transplantation can improve disease-free survival, overall survival and reduce post-transplant mortality. Monitoring of minimal residual disease (MRD) by flow cytometry and PCR-based molecular biology techniques is a widely reliable and practicable method. OBJECTIVE: To dynamically monitor the MRD level of acute lymphoblastic leukemia after peripheral blood haploidentical hematopoietic stem cell transplantation and to explore its implications for predicting early relapse. METHODS: A retrospective study was conducted in 53 patients with acute lymphoblastic leukemia who had underwgone peripheral blood haploidentical hematopoietic stem cell transplantation at the First Affiliated Hospital of Zhengzhou University from June 2011 to June 2017. The patients were followed up for postoperative 1, 3, 6, 12 months to observe the relation between MRD levels and relapse after transplantation. RESULTS AND CONCLUSION: (1) The disease-free survival rate of MRD positive group and MRD negative group were 20.0% and 65.8%, respectively; and the overall survival rates were 50.8% and 68.9% in the two groups, respectively. There were significant differences between two groups. (2) Among 16 MRD positive patients accepting clinical intervention after transplantation, 4 patients presented with MRD negative and had no recurrence. (3) Eleven hematologic recurrence patients were given tyrosine kinase inhibitor-targeted therapy, chemotherapy, donor lymphocytes Infusion and secondary transplantation, but they eventually died. The median time from the discovery of MRD positive to hematologic recurrence was 100 (7-190) days, and during this period. Clinical intervention was confirmed to extend the recurrence time. In this study, one case refused clinical intervention, and eventually died of recurrence. Our findings indicate that dynamic monitoring of the MRD level in acute lymphoblastic leukemia patients after peripheral blood haploidentical transplantation can predict recurrence, by which the patients can be given early intervention to reduce the risk of recurrence and improve disease-free survival and overall survival. 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 brain-derived neurotrophic factor-modified human amniotic membrane-derived mesenchymal stem cell transplantation on learning and memory abilities of Alzheimer's disease rats Gao Ming-long, Shi Shao-xia, Zhang Kun, Zhang Ying-dong, Li Na, Yu Ming, Wang Yong-liang 2018, 22 (9):  1419-1424.  doi: 10.3969/j.issn.2095-4344.0474 Abstract ( 335 )   PDF (6175KB) ( 202 )   Save BACKGROUND: A variety of stem cells have been found to be effective in the treatment of Alzheimer's disease in rats. However, few reports have been reported on the treatment of Alzheimer's disease rats with brain-derived neurotrophic factor (BDNF)-modified human amniotic membrane-derived mesenchymal stem cells. OBJECTIVE: To investigate the effects of BDNF-modified human amniotic membrane-derived mesenchymal stem cell transplantation on the learning and memory abilities of Alzheimer's disease rats. METHODS: Forty-eight Sprague-Dawley rats were divided into control group (no treatment), model group (Alzheimer's disease model), stem cell transplantation group (human amniotic membrane-derived mesenchymal stem cell transplantation+Alzheimer's disease model) and BDNF-modified stem cell transplantation group (BDNF-modified human amniotic membrane-derived mesenchymal stem cell transplantation+Alzheimer's disease model), 12 rats in each group. Learning and memory of model rats were determined in a trisection radiation maze and immunohistochemical staining was used to determine the number of p75 positive neurons at 2 weeks after cell transplantation. RESULTS AND CONCLUSION: The number of p75 positive neurons in the bevel zone and medial septal nucleus was ranked as follows: the model group < the stem cell transplantation group < the BDNF-modified stem cell transplantation group < the control group, and there were significant differences among groups (P < 0.05). The learning and memory abilities of the rats were ranked as follows: the model group < the stem cell transplantation group < the BDNF-modified stem cell transplantation group < the control group, and there were significant differences among groups (P < 0.05). In the BDNF-modified stem cell transplantation group, the number of learnings was negatively correlated with the number of p75 NGFR-positive neurons (P < 0.05), while the memory capacity was positively correlated with the number of p75 NGFR-positive neurons (P < 0.05). These findings reveal that human amniotic membrane-derived mesenchymal stem cell transplantation can improve learning and memory abilities of Alzheimer's disease rats, and BDNF-modified human amniotic membrane-derived mesenchymal stem cells can further improve this therapeutic effect. 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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Transplantation of lentiviral-transfected endothelial progenitor cells for pulmonary hypertension and the interventional effect of astragalosides Zhou Xiao-xiong, Wei Wei-chao, Sun Ce, Ye Tao-chun, Wang Song, Qing Li-jin, Wu Hui, Xian Shao-xiang 2018, 22 (9):  1425-1431.  doi: 10.3969/j.issn.2095-4344.0475 Abstract ( 387 )   PDF (6499KB) ( 209 )   Save BACKGROUND: Activation of CD40 pathway negatively regulates the therapeutic effect of endothelial progenitor cells (EPCs), while inhibition of this pathway can enhance the biological function of the cells. OBJECTIVE: To compare the therapeutic effects of CD40-silenced EPCs (EPCs shRNA-CD40) and conventional EPCs transplantation in an animal model of pulmonary hypertension, and to explore the interventional effect of astragalosides on EPCs transplantation in the treatment of pulmonary hypertension. METHODS: Ninety SD rats were randomly divided into five groups: normal control group (n=24), model group (n=24), lentivirus transfection group (n=18), conventional transplantation group (n=18) and astragaloside group (n=6). Except the normal control group, the remaining four groups were given monocrotaline to induce pulmonary hypertension. Rats in the lentivirus transfection and conventional transplantation groups were given intravenous injection of Lv-shRNA-CD40-transfected EPCs and conventional EPCs respectively at 7, 14, 21 days after modeling (n=6 at each time point). Rats in the astragaloside group were given daily intraperitoneal injection of 80 mg/(kg•d) astragaloside within 1-21 days after modeling, and then Lv-shRNA-CD40-transfected EPCs were intravenously injected at 21 days after modeling. Hemodynamics, plasma endothelin-1 level and right ventricular hypertrophy index were detected at 28 days after modeling. RESULTS AND CONCLUSION: (1) After modeling, right ventricular pressure, mean pulmonary arterial pressure and right ventricular hypertrophy index were all increased compared with the normal control group (P < 0.05), while these indices were then decreased significantly after EPCs transplantation (P < 0.05). With the increasing of transplantation time, there was an increasing trend in the right ventricular pressure, mean pulmonary arterial pressure and right ventricular hypertrophy index in the two EPCs transplantation groups, but this trend was not remarkable in the lentivirus transfection group. (2) After modeling, the level of endothelin-1 was increased significantly compared with the normal control group (P < 0.05), and then decreased after EPCs transplantation (P < 0.05). The level of endothelin-1 in the lentivirus transfection group was significantly lower than that in the conventional transplantation group at the same time point (P < 0.05). (3) A significant improvement in hemodynamics, plasma endothelin-1 level and right ventricular hypertrophy index was observed in the astragaloside group as compared with the lentivirus transfection group (P < 0.05). Given the above findings, CD40-silenced EPCs transplantation is more effective and durable than the conventional transplantation in the treatment of pulmonary hypertension, and moreover, astragaloside can enhance the therapeutic effect. 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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High-proportion differentiation of neural stem cells into neurons and formation of neural networks induced by active biomaterial scaffolds in vitro Li Ying, Zhang Ai-feng, Gao Yu-dan, Zhao Wen, Duan Hong-mei, Hao Peng, Shang Jun-kui, Yang Zhao-yang, Li Xiao-guang 2018, 22 (9):  1432-1437.  doi: 10.3969/j.issn.2095-4344.0476 Abstract ( 373 )   PDF (5649KB) ( 195 )   Save BACKGROUND: Either good biocompatibility and biological activity of active biological materials or the potential of multidirectional differentiation of neural stem cells has great application prospect and value. OBJECTIVE: To investigate the effect of neurotrophic factor 3-chitosan active biomaterial scaffolds on the differentiation of neural stem cells and the expression of key proteins of the neurotrophic factor 3 signal pathway in vitro. METHODS: The neural stem cells were extracted and purified, and then divided into pure culture medium group, soluble neurotrophic factor 3 group, pure chitosan group, and neurotrophic factor 3-chitosan group for differentiation induction. The expression of TrkC, Akt / p-Akt and Erk/p-Erk in the neurotrophic factor 3 signaling pathway was detected by western blot after 6 hours of induction. After 7 days of induction, differentiation of neural stem cells was observed by immunocytochemistry of MAP2, MBP, and GFAP. After 14 days of induction, formation of neural network induced by neurotrophic factor 3-chitosan active biomaterials was observed by immunocytochemistry of MAP2, Synapsin-1, and PSD95. RESULTS AND CONCLUSION: The neurotrophic factor 3-chitosan group induced a high proportion of neural stem cells differentiated into neurons, with a ratio of 73.8%, which was significantly higher than that in the other three groups. Meanwhile, the proportion of cells differentiated into glial cells waslower than that in the other three groups. The expression of key proteins TrkC, p-Akt and p-Erk in the neurotrophic factor 3-chitosan group was higher than that in the other three groups. Meanwhile, neurotrophic factor 3-chitosan could induce the in vitro differentiation of neural stem cells to form neural network. 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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Efficacy of autologous peripheral blood mononuclear cells in treatment of high radial nerve injury Yue Feng-wen, Liu Li-ping, Sun Guang-feng, Wu Xiang-kui, Wei Zai-rong, Wang Da-li 2018, 22 (9):  1438-1442.  doi: 10.3969/j.issn.2095-4344.0457 Abstract ( 361 )   PDF (1131KB) ( 412 )   Save BACKGROUND: In recent years, neural stem cells, adipose tissue derived mesenchymal stem cells, muscle derived stem cells and induced pluripotent stem cells have been applied in the treatment of peripheral nerve injuries. However, the treatment of peripheral nerve injury by autologous peripheral blood mononuclear cells has rarely been reported. OBJECTIVE: To investigate the efficacy of autologous peripheral blood mononuclear cells in the treatment of high radial nerve injury.  METHODS: From April 2011 to September 2015, 12 cases of radial nerve injury in the middle arm were treated. Preoperatively peripheral blood mononuclear cells were mobilized, and then 15 mL of mononuclear cell suspension was prepared on the operation day. Radial nerves scheduled for anastomosis were surgically explored and subjected to end-end anastomosis using outer membrane suturing under microscope. The anastomotic site of the nerve was enveloped with gelatin sponge soaked with 5 mL of autologous peripheral blood mononuclear cell suspension. The remaining 10mL of cell suspension was used for a multi-point injection into the local muscles, 0.5 mL at each point. Thereafter, the deep fascia and the incision were sutured in sequence. Postoperative antibiotics were used to prevent infection for 48 hours, and upper limb immobilization lasted for 4 weeks. Performance of rehabilitation exercise was guided. During the follow-up, wrist dorsal extension and muscle strength of extensor carpi ulnaris and extensor digitorum communis were detected to evaluate the therapeutic efficacy. RESULTS AND CONCLUSION: All the patients were followed up for 15 to 36 months, with an average of 17 months. Efficacy was excellent in 9 cases, good in 2 cases, fair in 1 case and poor in 0 case. The excellent and good rate was 92%. The wrist dorsal extension could achieve the functional needs, and the thumb dorsal extension and finger extension basically met the functional requirements. It is suggested that autologous peripheral blood mononuclear cell transplantation can achieve good outcomes in the treatment of high radial nerve 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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Combined use of 1,25-dihydroxyvitamin D3 and dexamethasone promotes osteogenic differentiation of synovial fluid mesenchymal stem cells from human temporomandibular joint Liu Wen-jing, Sun Yang-peng, Zheng You-hua, Zhang Zhi-guang 2018, 22 (9):  1443-1449.  doi: 10.3969/j.issn.2095-4344.0477 Abstract ( 386 )   PDF (5750KB) ( 216 )   Save BACKGROUND: Synovial fluid mesenchymal stem cells can be amplified rapidly in vitro and collected by a minimally invasive method. Recent studies have suggested that synovial fluid mesenchymal stem cells have become an important kind of seed cells for bone tissue engineering. Osteogenic differentiation is required to be optimized prior to the application of synovial fluid mesenchymal stem cells in the bone regeneration. OBJECTIVE: To describe a protocol to generate osteoblast-lineage cells from human synovial fluid mesenchymal stem cells of the temporomandibular joint using a cocktail that includes glutamax, dexamethasone, β-glycerophosphate, vitamin C, 1,25-dihydroxyvitamin D3, and to investigate the effect of 1,25-dihydroxyvitamin D3 and dexamethasone on the osteogenic capacity of synovial fluid mesenchymal stem cells. METHODS: Synovial fluid mesenchymal stem cells from the human temporomandibular joint were expanded in vitro and cultured in different osteogenic induction media. The mineralization capacity of osteogenic differentiation was evaluated by alizarin red and Von kossa staining. And the osteogenic markers, including ALP, RUNX2 and OCN, were assessed by reverse transcription-PCR. RESULTS AND CONCLUSION: The mineralization formation increased greatly in the medium with 100 nmol/L dexamethasone and 10 nmol/L 1,25-dihydroxyvitamin D3. The expression of ALP, RUNX2 and OCN was enhanced distinctly in the 1,25-dihydroxyvitamin D3-induced differentiated cells. These findings reveal that appropriate concentration of 1,25-dihydroxyvitamin D3 and dexamethasone can be ideal ingredients to induce the osteogenic differentiation of human synovial fluid mesenchymal stem cells of the temporomandibular joint. Thus, this effective condition can be used to induce the osteogenic differentiation of synovial fluid mesenchymal stem cells for the bone regeneration in the temporomandibular joint. 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 sustained hypoxia on proliferation of mouse embryonic fibroblasts and preparation of feeder layers Wei Han-qing, Pei Yi-jin, Wang Dan-dan, Jiang Yang, Wang Chun, Li Hong-mei 2018, 22 (9):  1450-1456.  doi: 10.3969/j.issn.2095-4344.0478 Abstract ( 504 )   PDF (5773KB) ( 224 )   Save BACKGROUND: In traditional culture systems for embryonic stem cells, feeder cell preparation and embryonic stem cell culture are mostly performed under normoxic conditions. Changes in oxygen culture conditions are likely to influence feeder cells, thereby altering the growth characteristics or differentiation ability of embryonic stem cells, but there is still no relevant systematic report until now. OBJECTIVE: To investigate the effects of sustained hypoxia culture on the pluripotency of mouse embryonic stem cells cultured on mouse embryonic fibroblast feeder layers. METHODS: Primary mouse embryonic fibroblasts were persistently subcultured under normoxia (20% O2) and hypoxia (5% O2) conditions. Cell proliferation was measured for drawing growth curve. Reactive oxygen species level and mitochondria membrane potential of the feeder cells were detected respectively. Mouse embryonic stem cells were divided into two groups: normoxia group (plated on mouse embryonic fibroblast feeder layers under 20% O2), and hypoxia group (plated on mouse embryonic fibroblast feeder layers under 5% O2). The cell morphology was observed and the pluripotency of embryonic stem cells were detected by measurement of Oct4 and Sox2 expressions. Hypoxia inducible factor-1α mRNA expression was also tested in the four groups. RESULTS AND CONCLUSION: As compared to the normoxia group, mouse embryonic fibroblasts in the hypoxia group proliferated faster, reactive oxygen species significantly declined, and the mitochondria membrane potential level increased significantly (P < 0.05). Embryonic stem cells were positive for alkaline phosphatase, and highly expressed Oct4 and Sox2 mRNA. Much more median- or small-sized colonies formed in the hypoxia group than the normoxia group (P < 0.05). The mRNA expression of hypoxia inducible factor-1α in embryonic stem cells had a significant difference between the hypoxia and normoxia groups (P < 0.05). These findings indicate that a sustained hypoxia environment can significantly promote the viability of mouse embryonic fibroblasts as feeder layers and maintain the pluripotency of embryonic stem cells under 5% O2. 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 TAZ, a transcriptional coactivator, on human dental pulp stem cell proliferation in vitro Zhang Wen, Luo Shi-wei, Gu Ai-ling, He Wei 2018, 22 (9):  1457-1462.  doi: 10.3969/j.issn.2095-4344.0479 Abstract ( 290 )   PDF (3979KB) ( 181 )   Save BACKGROUND: How to improve the proliferation of human dental pulp stem cells (hDPSCs) in vitro is of great importance in regenerative medicine. OBJECTIVE: To investigate the effect of TAZ, a transcriptional coactivator, on the proliferation of hDPSCs and its potential mechanisms.  METHODS: hDPSCs were isolated from human dental pulps and cultured in vitro. The expression of TAZ in hDPSCs was detected by immunofluorescence method. Then, we knocked down the expression of TAZ in hDPSCs, and the proliferation of hDPSCs was measured by MTT and BrdU kit analysis respectively. The levels of CTGF and Cry61, which are the downstream target genes of TAZ, were detected by RT-qPCR and western blot assay. We also investigated the effect of TAZ on the levels of transforming growth factor-β (TGF-β) signaling proteins, Smad3 and Smad 4 by using western blot. RESULTS AND CONCLUSION: TAZ protein was expressed in hDPSCs. After the transfection by siTAZ for 24 hours, the mRNA and protein levels of TAZ were both significantly decreased. After the transfection by siTAZ for 24 and 48 hours, the proliferation of hDPSCs was obviously inhibited. Meanwhile, the mRNA and protein levels of CTGF and Cry61 were decreased by siTAZ transfection. The further research showed that TAZ silence also inhibited the expression of Smad3 and Smad 4, which belonged to the TGF-β signaling pathway. To conclude, TAZ may modulate the proliferation of hDPSCs through regulating the expression of CTGF and Cry61, and TGF-β-dependent signaling pathways may be involved in this regulation.  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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Clinical treatment of heart failure by adult stem cells: existing problems and prospects Lu Meng-xiao, Zheng Yuan-yuan, Liu Yang, Lu Jian-wei, Li Peng, Wu Ming-yuan 2018, 22 (9):  1463-1469.  doi: 10.3969/j.issn.2095-4344.0480 Abstract ( 403 )   PDF (1189KB) ( 256 )   Save BACKGROUND: The prevalence of heart failure is increasing yearly. Although traditional treatment methods have made great progress in alleviating the progression of heart failure, cardiac tissue injury cannot be thoroughly cured. Cell therapy, however, has the potential to completely cure it through myocardial regeneration.  OBJECTIVE: To review the current progress in adult stem cells from different sources in the treatment of heart failure. METHODS: A computer-based retrieval of PubMed and CNKI databases was performed in order to search relevant articles published from 2001 till now, using the keywords of “heart failure, adult stem cells, skeletal myoblasts, cardiac stem cells, mesenchymal stem cells. After removal of repetitive or irrelevant articles, 61 articles were finally reviewed. RESULTS AND CONCLUSION: Currently used adult stem cells for treating heart failure mainly include skeletal myoblasts, bone marrow mesenchymal stem cells, adipose-derived mesenchymal stem cells, human umbilical cord blood mesenchymal stem cells and cardiac stem cells. Most clinical results have shown that adult stem cells have a good effect in the treatment of heart failure, and cause few adverse reactions. The mechanism of adult stem cells in the treatment of heart failure may be related to post-transplantation angiogenesis, paracrine mechanisms, and cell fusion, but the choices of specific cell lines, dose, route of administration and treatment frequency as well as the precise mechanism of action still need further studies. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Human umbilical cord-derived mesenchymal stem cells in the treatment of myocardial infarction: existing problems, effects and prospects Wang Yuan-fei, Li Ya-xiong, Zhang Ya-yong, Jiang Li-hong 2018, 22 (9):  1470-1476.  doi: 10.3969/j.issn.2095-4344.0481 Abstract ( 513 )   PDF (952KB) ( 274 )   Save BACKGROUND: Human umbilical cord-derived mesenchymal stem cells, because of convenient acquisition, will not bring pain and adverse effects to the fetus and their parturient. The success rate of culture is high. It has good application prospects in the treatment of myocardial infarction. OBJECTIVE: To review the new progress of human umbilical cord-derived mesenchymal stem cells transplantation in the treatment of myocardial infarction. METHODS: A computer-based online search of PubMed and Wanfang databases was performed to retrieve the related articles published from 1991 to 2017. We reviewed the initial data and the quotations from each document. Finally, we included randomized controlled animal experiments or clinical studies concerning human umbilical cord-derived mesenchymal stem cells for treatment of myocardial infarction. RESULTS AND CONCLUSION: Human umbilical cord-derived mesenchymal stem cells will become a new alternative source of cells in the treatment of myocardial infarction, with extremely broad prospects. However, there is no mature conclusion about the convenient route, optimal number of transplanted cells, optimal timing, differentiation, homing and evaluation after transplantation, which limits the clinical application of these 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