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03 June 2016, Volume 20 Issue 23 Previous Issue    Next Issue

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Inhibitory effect of miR-34a on lung cancer stem cells via Notch1 signaling pathway Han Ji-chang, Zhang Yi-jie, Li Hong-bing, Yang Cun-bao, Ma Chao-nan, Qi Guan-bin 2016, 20 (23):  3349-3356.  doi: 10.3969/j.issn.2095-4344.2016.23.001 Abstract ( 306 )   PDF (1345KB) ( 247 )   Save BACKGROUND: It has been proved that miR-34a plays an inhibitory role in the growth of lung cancer stem cells, but the underlying mechanism remains unclear. OBJECTIVE: To explore the inhibitory effect of miR-34a on lung cancer stem cells and the underlying mechanism. METHODS: The CD133+ lung cancer stem cells were separated from lung cancer A549 cell lines using magnetic activated cell sorting method. And miR-34a-overexpressing CD133+ lung cancer stem cells were established by liposome transfection technology. Besides, the targeted relationship between miR-34a and Notch1 was analyzed by the dual-luciferase reporter. Afterwards, Notch1 silencing was performed by gene knockout, and its effect on lung cancer stem cells was determined. RESULTS AND CONCLUSION: After sorted and detected by immunomagetic selection and flow cytometry assay respectively, a high rate of CD133+ lung cancer stem cell was obtained. And qRT-PCR detected that the expression level of miR-34a in CD133+ lung cancer stem cells was significantly lower than that in CD133- lung cancer stem cells. Moreover, miR-34a-overexpressing CD133+ lung cancer stem cells were successfully constructed and miR-34a significantly inhibited proliferation and induced apoptosis of lung cancer stem cells. Dual-luciferase reporter assay indicated that Notch1 mRNA was a target of miR-34a. In addition, Notch1 silencing obviously inhibited proliferation and induced apoptosis of lung cancer stem cells. These findings suggest that miR-34a can inhibite lung cancer stem cells via the Notch1 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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Peroxisome proliferator-activated receptor gamma promotes exogenous mesenchymal stem cells to express connexin 43: its role and mechanism Yan Ping, Hou Jing-ying, Zheng Shao-xin, Long Hui-bao, Zhou Chang-qing, Guo Tian-zhu, Wu Quan-hua, Zhong Ting-ting, Wang Tong 2016, 20 (23):  3357-3365.  doi: 10.3969/j.issn.2095-4344.2016.23.002 Abstract ( 279 )   PDF (5805KB) ( 245 )   Save BACKGROUND: Piglitazone, a peroxisome proliferator-activated receptor γ (PPAR-γ) agonist, has been demonstrated to promote survival and cardiac differentiation of exogenous bone marrow mesenchymal stem cells to improve cardiac function. In this study, we attempted to investigate whether pioglitazone could induce cardiac differentiation of endogenous bone marrow mesenchymal stem cells and improve cardiac function, and meanwhile, probed into the relevant mechanisms. OBJECTIVE: To compare the therapeutic efficacy of pioglitazone combined with bone marrow mesenchymal stem cell transplantation, pioglitazone alone and phosphate buffer solution (PBS) and to investigate the relevant mechanisms. METHODS: Thirty Sprague-Dawley rats with myocardial infarction induced by ligation of the left anterior descending coronary artery were randomized into combined group (combination of bone marrow mesenchymal stem cells and pioglitazone), pioglitazone group and PBS group. Two weeks later, PKH26-labeled bone marrow mesenchymal stem cells in PBS or PBS alone were injected into the local infarct zone in the combined group and the other two groups, respectively. Pioglitazone (3 mg/kg/d) was given by the oral gavage in the combined and pioglitazone groups for continuous 2 weeks after cells transplantation. At 2 weeks after treatment, cardiac functions were evaluated. In addition, expressions of PPAR-γ, connexin 43 and relative factors in transforming growth factor-β1/SMAD signaling pathway were examined in different areas of the left ventricle from each harvested heart. RESULTS AND CONCLUSION: There were no differences in the baseline parameters of cardiac function between the two groups. Two weeks after treatment, left ventricular end-diastolic diameter, left ventricular end-systolic diameter and left ventricular ejection fraction were significantly improved in the combined group compared with the other two groups; the expression of PPAR-γ was significantly increased in different zones of the left ventricle in the combined and pioglitazone groups. In the combined group, there was a significantly higher expression of connexin 43, and the levels of transforming growth factor-β1, SMAD2 and SMAD3 were obviously attenuated in the infarct and marginal zones. However, no differences were found in the above determinants between the pioglitazone and PBS groups. To conclude, pioglitazone cannot induce the differentiation and proliferation of endogenous bone marrow mesenchymal stem cells, but pioglitazone combined with exogenous bone marrow mesenchymal stem cells can improve cardiac function post myocardial infarction. In this process, PPAR-γ might promote the connexin 43 expression in exogenous bone marrow mesenchymal stem cells via the blockade of transforming growth factor-β1/SMAD 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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Properties of human bone marrow mesenchymal stem cells in the early phase of adipogenic differentiation in different culture systems Tian Jian-jian, Chi Ying, Song Bao-quan, Du Wen-jing, Han Zhi-bo, Han Zhong-chao 2016, 20 (23):  3366-3373.  doi: 10.3969/j.issn.2095-4344.2016.23.003 Abstract ( 328 )   PDF (6130KB) ( 312 )   Save BACKGROUND: There are various methods to induce adipogenic differentiation of bone marrow mesenchymal stem cells, and the main component for adipogenic induction is indomethacin or rosiglitazone. However, there is a lack of comparative study on the induction efficiency and mechanism among these methods. OBJECTIVE: To compare the adipogenic responses of human bone marrow mesenchymal stem cells to different induction methods, and to analyze the mechanism underlying different induction efficiency. METHODS: After isolation and purification, the adipogenic abilities of human bone marrow mesenchymal stem cells in three different culture systems were compared by oil red O staining and lipogenic gene assay. At 0, 1, 3 and 7 days of adipogenensis, mRNA expressions of PPARγ, C/EBPα, Adiponectin and Leptin were detected. At 7 days of adipogenensis, protein expressions of PPARγ and C/EBPβ were detected by western blot assay, and effects of DIMI versus DIMR on phosphorylation of PPARγ at Ser273 were compared. RESULTS AND CONCLUSION: Findings from oil red O staining and real-time PCR showed that DIMR significantly induced adipogenic differentiation of bone marrow mesenchymal stem cells compared with DIM and DIMI at 7 days of induction. Western blot showed that the protein expressions of PPARγ and C/EBPβ in the DIMI group were significantly higher than those in the DIMR and DIM at 7days of induction. In addition, the ratio of PPARγ phosphorylation at Ser273 was lower in the DIMR group than the DIMI group. To conclude, DIMR has the most potential to induce early adipogenesis of human bone marrow mesenchymal stem cells by weakening the phosphorylation of PPARγ-Ser273.   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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CD34, a cancer stem cell marker, in nasopharyngeal carcinoma cell lines Zhang Jun-hong, Li Qing, Wang Chun-hua, Wu Xiu-yun, Lu Xin-ge, Wang Li-fei, Yang Lei-lei,Wang Qiu-hong 2016, 20 (23):  3374-3379.  doi: 10.3969/j.issn.2095-4344.2016.23.004 Abstract ( 377 )   PDF (3845KB) ( 235 )   Save BACKGROUND: Previous research have confirmed that CD34 is closely related to oncogenesis, progress, recurrence, metastasis and drug-resistance of various cancers, but its role in nasopharyngeal carcinoma remains unclear. OBJECTIVE: To sort cells positive and negative for CD34 in nasopharyngeal carcinoma cell lines and to detect cell proliferation and migration. METHODS: Expressions of CD34 in nasopharyngeal carcinoma cell lines 5-8F, 6-10B, CNE1 and CNE2 were detected by flow cytometry. And CD34+ and CD34- cells were sorted based on cell surface markers for purity identification. Afterwards, proliferation and migration of CD34+ and CD34- cells were detected by MTT assay, colony-formation assay and scratch assay. RESULTS AND CONCLUSION: All four nasopharyngeal carcinoma cell lines expressed CD34 in 0.1%-0.2%, and the level of CD34 was closely related to the cell growth density. The purity of CD34+ cell was more than 98% in the sorted CD34+ cell populations, but no CD34+ cells were found in the sorted CD34- cell populations. At 1, 3, 5 and 7 days the proliferation rate of CD34+ cell, populations was significantly higher than that of CD34- cells (P < 0.05). Consistently, the colony-formation efficiency of CD34+ cell was significantly higher than that of CD34- cells (P < 0.05). Moreover, CD34+ cells migrated significantly faster than CD34- cells by scratch assay (P < 0.05). In conclusion, CD34+ cells cultured in vitro display higher proliferation and migration capacities, indicating that CD34+ cells have the potential of nasopharyngeal carcinoma 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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Differentiation and culture of tumor stem cells in human primary gastric cancer cells Wang Ying 2016, 20 (23):  3380-3385.  doi: 10.3969/j.issn.2095-4344.2016.23.005 Abstract ( 347 )   PDF (3319KB) ( 220 )   Save BACKGROUND: There is a close relationship between tumor stem cells and tumor occurrence and recurrence, but there are still some disputes in the presence of tumor stem cells in all tumors. OBJECTIVE: To investigate the differentiation and culture of tumor stem cells in human primary gastric cancer cells. METHODS: Primary gastric cancer cells isolated from fresh gastric cancer tissues were stained with hematoxylin-eosin and identified by immunohistochemical detection of carcinoembryonic antigen. The CD44 expression of the cells was detected using immunofluorescence method. Magnetic activated cell sorting was used to isolate CD44+ gastric cancer cells that were then seeded subcutaneously behind the armpit of mice. Growth of implanted tumor cells was observed. RESULTS AND CONCLUSION: Human primary gastric cancer cells were isolated in serum-free medium. Compared with the routine culture group, the number of CD44+ cells (P < 0.05) and the tumor volume were significantly increased in the spheroid culture group. Furthermore, at 90 days after transplantation, the tumor volume of mice in spheroid culture group was significantly higher than that in the routine culture group. These experimental findings indicate that gastric cancer cells with certain tumorigenicity can be successfully isolated from gastric cancer cells using serum-free culture method and magnetic activated cell sorting method.   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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Recombinant adenovirus-mediated hypoxia inducible factor 1alpha mutant promotes proliferation of adipose-derived mesenchymal stem cells Zhang Xin, Zhou Lin, Zhang Xiao-lei 2016, 20 (23):  3386-3393.  doi: 10.3969/j.issn.2095-4344.2016.23.006 Abstract ( 255 )   PDF (5116KB) ( 238 )   Save BACKGROUND: To improve the survival rate of transplanted tissue, most scholars focus on cell therapy, particularly cell-assisted fat grafting. OBJECTIVE: To analyze the effect of recombinant adenovirus-mediated hypoxia inducible factor 1alpha (HIF1α) mutant on survival rate of transplanted fat particles through transfection of adipose-derived mesenchymal stem cells. METHODS: Recombinant adenovirus-mediated triple-mutant HIF1α was inserted into an adenovirus pAdEasy-1 system, followed by viral packaging and titer determination. Human adipose-derived mesenchymal stem cells were cultured, passaged and identified, and subsequently transfected with three kinds of viruses and blank vector (experimental group with transfection of the triple mutant of HIF1α; positive control group; negative control group; blank control group). Transfection efficiency was determined using enhanced green fluorescent protein labeling. Additionally, MTT assay was used to detect cell proliferation. RESULTS AND CONCLUSION: The recombinant adenovirus was successfully constructed and packaged in line with transfection requirements. Moreover, adipose-derived mesenchymal stem cells were successfully identified by adipogenic, osteogenic and chondrogenic induction and could be used as seed cells for subsequent experiments. RT-PCR results showed that HIF1α mRNA expression in the experimental group and positive control group was significantly higher than that in the other two groups(P < 0.05). Western blot analysis showed that the relative absorbance value in the experimental group was significantly higher than that in the other three groups (P < 0.05). A significant increase in the cell proliferation was found in the experimental group, significantly different from the other three groups (P < 0.05). Therefore, our findings indicate that transfection of adenovirus-mediated triple-mutant HIF1α not only can sustain the expression of target protein in transfected adipose-derived mesenchymal stem cells under normoxic conditions, but also can promote the proliferation of transfected 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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Induced differentiation of human embryonic stem cells into endothelial cells Shi Yang, Shen Jun-sheng, Wu Tao-tao, Li Xiao-fei 2016, 20 (23):  3394-3399.  doi: 10.3969/j.issn.2095-4344.2016.23.007 Abstract ( 292 )   PDF (3542KB) ( 283 )   Save BACKGROUND: Human embryonic stem cells exhibit self-renewal and multi-differentiation potential, and can differentiate into endothelial cells under certain induction conditions. OBJECTIVE: To explore induced conditions of the human embryonic stem cells differentiating into endothelial cells and to investigate the effect of vascular endothelial growth factors on the endothelial differentiation of human embryonic stem cells. METHODS: After resuscitation, passage 40 human embryonic stem cell lines H9 were subjected to suspension culture to prepare embryos, and after 5-day culture, these cells were cultured in attachment medium to differentiate into embryoid bodies, followed by induction with 50 µg/L vascular endothelial growth factors. Passage 2 and 15 embryonic stem cells after induced differentiation were taken for Dil-Ac-LDL uptake test and immunohistochemical staining, respectively. RESULTS AND CONCLUSION: After 1-day culture, cord-like or polygonal monolayer cells around embryoid bodies showed bud-like and radial growth with a relative rapid speed merging into surrounding colonies; at 2-3 days, the number of suspension cells increased further, but the small-round cells in the center began to die; at 5 days, embryoid bodies started to passage, and aggregated cells exhibited typical paving stone-like appearance. Moreover, some human embryonic cells after induced differentiation could actively take up fluorescent labeled LDL, and red fluorescent particles appeared. Additionally, passage 15 embryonic stem cells after induced differentiation could express CD31 and FLK-1. These findings suggest that human embryonic stem cells induced by vascular endothelial growth factors can differentiate into endothelial 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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PTEN-silenced bone marrow mesenchymal stem cell transplantation for acute myocardial infarction Xu Hui-min, Liu Yu-ru, Wan Qi-lin, Li Yan-ming, Wu Chang-wei 2016, 20 (23):  3400-3406.  doi: 10.3969/j.issn.2095-4344.2016.23.008 Abstract ( 348 )   PDF (4758KB) ( 558 )   Save BACKGROUND: Bone marrow mesenchymal stem cell transplantation for myocardial infarction becomes popularized in recent years, but transplanted cells cannot survive and proliferate under early inflammatory reaction or local ischemia/hypoxia microenvironment, eventually hampering the therapeutic outcomes. OBJECTIVE: To investigate the therapeutic effect of PTEN-silenced bone marrow mesenchymal stem cells on acute myocardial infarction. METHODS: (1) Bone marrow mesenchymal stem cells from Sprague-Dawley rats were randomly assigned to receive no treatment, NCsiRNA transfection using Lipofectamin2000 or PTEN siRNA transfection using Lipofectamin2000. Cell growth curves were described using MTT method to detect cell cycle using flow cytometry. (2) Thirty Sprague-Dawley rats were selected to prepare myocardial infarction models that were randomized into three groups (n=10 per group): blank control, negative control and RNAi group. Six hours after modeling, bone marrow mesenchymal stem cells transfected with nothing, NCsiRNA and PTEN siRNA were respectively injected into the infarcted center of the left ventricular anterior wall in these three rat groups. After 4 weeks, all rats were subjected to cardiac function detection using echocardiography, and the survival and proliferation of bone marrow mesenchymal stem cells in the rats were observed by fluorescence microscopy. RESULTS AND CONCLUSION: Compared with the other two groups, a significant increase in the absorbance values at different culture time, the proportion of cells in S+G2 phase, and the number of bone marrow mesenchymal stem cells in the myocardial tissue was found in the RNAi group (all P < 0.05). Additionally, the left ventricular ejection fraction and left ventricular shortening fraction were significantly reduced in the RNAi group than the blank control and negative control groups at 4 weeks after cell transplantation (P < 0.05). Both in vivo and in vitro experimental findings showed that PTEN silencing could effectively improve cell survival and proliferation in the infarcted myocardium. Moreover, in the in vivo experiment, an overt improvement in rat’s cardiac function was achieved.    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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Bioinformatics analysis of differentially expressed oncogenes in liver cancer stem cells Li Shu-na, Song Na-na 2016, 20 (23):  3407-3412.  doi: 10.3969/j.issn.2095-4344.2016.23.009 Abstract ( 288 )   PDF (884KB) ( 260 )   Save BACKGROUND: Studies have suggested that oncogene expression in liver cancer stem cells has a certain relationship with the occurrence and development of liver cancer, but there is still a lack of research on bioinformatics and mechanisms. OBJECTIVE: To identify differentially expressed genes in liver cancer stem cells and to analyze these genes by bioinformatics. METHODS: The human hepatoma cell line HepG2 was the cell tool of liver cancer stem cells to prepare total RNA, and fluorescent labeling experiment was conducted. Using gene chips hybridization, mRNA expression profiles were obtained and were screened, and then differentially expressed mRNAs were obtained, GO and Pathway annotations were analyzed using bioinformatics. RESULTS AND CONCLUSION: The detection rate was 73.21% for hybridization experiment, indicating the hybridization experiment is successful. In this study, a total of 38 342 mRNA were found, and after further analysis, 1 236 differentially expressed genes were screened (P < 0.05, fold change ≥ 2), including 599 up-regulated and 637 down-regulated genes, respectively (P < 0.05). Biological functions of differentially expressed genes were mainly involved in the histone H4 acetylation, cell mitosis and proliferation, synthesis and decomposition of cell-associated proteins, chromosome segregation, cell differentiation and apoptosis, signal transduction, as well as nutrient transportation and transcription. Pathway annotations were mainly related to cytokine-mediated inflammatory signaling pathway, Wnt signaling pathway, Hedgehog and Notch signaling pathways, TGF-Beta, Jak-STAT and so on. These results demonstrate that differentially expressed genes in liver cancer stem cells are related to the occurrence of liver cancer, and may be involved in the regulation of signaling pathways, which provides a new target for the treatment of liver 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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Effect of bone marrow mesenchymal stem cells via intravenous transplantation on cardiac function in heart failure induced by cardiomyopathies Hou Yong-lan, Yang Dong-wei 2016, 20 (23):  3413-3418.  doi: 10.3969/j.issn.2095-4344.2016.23.010 Abstract ( 347 )   PDF (1084KB) ( 220 )   Save BACKGROUND: Bone marrow mesenchymal stem cell transplantation can effectively improve decreased cardiac function caused by heart failure, but there is a lack of research about the effect in bone marrow mesenchymal stem cell transplantation on cardiac function in heart failure induced by cardiomyopathies. OBJECTIVE: To explore the effect of bone marrow mesenchymal stem cell transplantation on cardiac function in patients with cardiomyopathies accompanied by heart failure. METHODS: Totally 40 Sprague-Dawley rats were enrolled, and bone marrow mesenchymal stem cells were isolated from 10 rats, and the remaining rats were equivalently randomized into normal, model and stem cell transplantation groups. Then rats in the model and stem cell transplantation groups were given intraperitoneal injection of hydrochloric acid doxorubicin to prepare cardiomyopathy-induced heart failure models. At 7 days after modeling, the stem cell transplantation group was treated with bone marrow mesenchymal stem cells through intravenous transplantation, and the model group was treated with equal amount of DMEM medium. Four weeks later, cardiac function of each rat was detected, and the cell survival and differentiation were observed by immunofluorescence method. RESULTS AND CONCLUSION: At 4 weeks after transplantation, compared with normal and stem cell transplantation groups, the left ventricular systolic pressure and maximum rise/fall rate of left ventricular pressure were significantly decreased, but the left ventricular end diastolic pressure significantly increased in the model group (P < 0.05). And there was no significant difference between the normal group and the stem cell transplantation group (P > 0.05). High and dense fluorescence intensity was observed in the host myocardium immediately after transplantation. Subsequently, the fluorescence intensity and density decreased at 4 weeks, but the cell migration could be found, and some cells expressed cardiac troponin T. These results show that intravenous transplantation of bone marrow mesenchymal stem cells can improve cardiac function in rats with heart failure due to cardiomyopathies. Besides, the transplanted cells can survive in the host, and differentiate into cardiomyocyte-like 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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Oligodendrocyte transplantation for acute spinal cord injury: insulin-like growth factor 1 induces bone marrow mesenchymal stem cells differentiating into oligodendrocytes Wei Fen, Xiao Xiu-lan 2016, 20 (23):  3419-3424.  doi: 10.3969/j.issn.2095-4344.2016.23.011 Abstract ( 344 )   PDF (1042KB) ( 249 )   Save BACKGROUND: Treatment and rehabilitation of spinal cord injury is a complicated problem, and the reconstruction and remyelination of neural reflex pathways are the essential process, during which oligodendrocytes play an important role in spinal cord injury repair. OBJECTIVE: To observe the effect of oligodendrocyte transplantation for acute spinal cord injury in rats. METHODS: Insulin-like growth factor 1 induced bone marrow mesenchymal stem cells differentiating into oligodendrocytes, and those oligodendrocytes were transplanted into rats with acute spinal cord injury as induced cell transplantation group. Simple normal saline and natural oligodendrocytes were transplanted into the rat injured spinal cord as control group and oligodendrocyte group, respectively. Rat behavioral changes were observed by inclined plane test and Basso-Beattie-Bresnahan (BBB) scores. Neurological recovery and survival of the transplanted cells was detected and observed using spinal evoked potential and immunohistochemical staining, respectively. RESULTS AND CONCLUSION: Compared with the control group, BBB scores and the critical elevation angle of the incline plane test significantly increased, latencies of spinal motor and sensory evoked potential were on the decline (P < 0.05), and there were no significant differences in above indicators between the two groups at 4 and 8 weeks after transplantation. Moreover, survived oligodendrocytes after transplantation could be found in the lesions of spinal cord in both two groups. In conclusion, insulin-like growth factor 1-induced bone marrow mesenchymal stem cells can differentiate into oligodendrocytes that exact an excellent role in acute spinal cord injury repair after transplantation, which achieve the equal clinical efficacy to the natural oligodendrocytes.   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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Tanshinone II combined with bone marrow mesenchymal stem cell transplantation promotes nerve regeneration following cerebral infarction Lu Yi-zhong, Li He-hua, Lu Yi-fan 2016, 20 (23):  3425-3431.  doi: 10.3969/j.issn.2095-4344.2016.23.012 Abstract ( 284 )   PDF (1186KB) ( 293 )   Save BACKGROUND: Studies have shown that tanshinone II can improve microcirculation, dilate cerebral blood vessels, increase cerebral blood flow, reduce infarct size, and improve brain function after cerebral metabolic disorder. OBJECTIVE: To investigate the effect of tanshinone II combined with bone marrow mesenchymal stem cell transplantation on nerve regeneration following cerebral infarction in rats. METHODS: Rat models of acute cerebral infarction were prepared using the thread occlusion method and then given tanshinone II combined with bone marrow mesenchymal stem cell transplantation (combined group), bone marrow mesenchymal stem cell transplantation (cell transplantation group), and no treatment (model group), respectively. Neuromotor function was assessed using Longa scores. Expression of brain-derived neurotrophic factor gene around the infarction region was detected using RT-PCR. Cell apoptosis was detected using TUNEL. Immunohistochemistry staining was used to determine peri-cortex Nogo-A and NgR protein expression at the infarction region. RESULTS AND CONCLUSION: Longa scores, apoptotic index, and expression of Nogo-A and NgR proteins exhibited significant differences among three groups (P < 0.05) as follows: combined group < cell transplantation group < model group. Conversely, the expression of brain-derived neurotrophic factor gene was highest in the combined group successively followed by the cell transplantation group and model group (P < 0.05). These data show that tanshinone II combined with bone marrow mesenchymal stem cell transplantation accelerate the recovery of neurologic function and promote nerve regeneration after cerebral infarction by incresing mRNA expression of 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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Human erythropoietin-modified human amniotic mesenchymal stem cells via subarachnoid transplantation promote neurologic recovery from brain injury Lv Gang, Zhang Hong-yuan 2016, 20 (23):  3432-3438.  doi: 10.3969/j.issn.2095-4344.2016.23.013 Abstract ( 416 )   PDF (1309KB) ( 266 )   Save BACKGROUND: Studies have shown that as a regulator of bone marrow function erythropoietin is a glycoprotein that controls the development of the central nervous system and has neurotrophic and neuroprotective potential. Therefore, transplantation of human amniotic mesenchymal stem cells genetically modified by human erythropoietin is a new choice for brain injury treatment. OBJECTIVE: To observe the effect of transplantation of human amniotic mesenchymal stem cells genetically modified by human erythropoietin on the functional recovery from brain injury in rats. METHODS: Eukaryotic expression plasmid pcDNA3.1 carrying erythropoietin was successfully constructed and transferred into amniotic mesenchymal stem cells cultured in vitro. Expression of erythropoietin was detected using western blot assay before and after transfection. Rat models of middle cerebral arterial occlusion was made and given transplantation of transfected amniotic mesenchymal stem cells via the tail vein (transfection group). Additionally, model and simple cell transplantation groups were set in a comparative study. RESULTS AND CONCLUSION: Findings from western blot detection showed that transfected cells could express human erythropoietin. Compared with the other groups, modified neurologic severity scores, growth-associated protein 43 and aquaporin 9 at mRNA and protein levels were all decreased significantly in the transfection group. Furthermore, the number of cells positive for CM-Dil was highest in the transfection group, followed by simple cell transplantation group, and lowest in the model group (all P < 0.05). Overall findings from this study show that human erythropoietin-modified human amniotic mesenchymal stem cell transplantation promotes neurologic recovery from brain injury through eliciting a reduction in growth-associated protein 43 and aquaporin 9 at mRNA and protein levels as well as inhibiting cell apoptosis.   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 Guiqi Dihuang Tang on the treatment of sensorineural deafness with cochlear stem cells Zhu Han-qing, Zhang Jun-feng, Chen You-dong, Xu Zhen 2016, 20 (23):  3439-3444.  doi: 10.3969/j.issn.2095-4344.2016.23.014 Abstract ( 337 )   PDF (1338KB) ( 194 )   Save BACKGROUND: Combining Chinese medicine with stem cell transplantation opens up a new avenue for the use of traditional Chinese medicine in the stem cell transplantation. It is expected to improve survival and differentiation of cochlear stem cells into hair cells through Chinese medicine interventions. OBJECTIVE: To observe the influence of Guiqi Dihuang Tang on the treatment of sensorineural deafness with cochlear stem cells. METHODS: Guinea pigs with sensorineural deafness were randomly assigned into three groups: combined group intervened with cochlear stem cell suspension with medicated serum, cell transplantation group with cochlear stem cell suspension, and blank control group with normal saline injection. At 7, 28 and 56 days after treatment, all guinea pigs were detected for auditory brainstem response and immunofluorescent observation. RESULTS AND CONCLUSION: Nestin positive cells and MyosinVIIA positive cells were observed in the combined and cell transplantation groups, and the number of positive cells was higher in the combined group than the cell transplantation group. Auditory brainstem response threshold of guinea pigs was decreased in the combined and cell transplantation groups, and the recovery of hearing was better in the former group. These findings indicate that the intervention of Guiqi Dihuang Tang can remarkably improve the survival rate of transplanted stem cells and the differentiation ratio of transplanted cells into hair 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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Intracoronary transplantation of umbilical cord blood stem cells reduces arrhythmia score and incidence of ventricular arrhythmia Wang Cheng, Guo Chang-lei, Li Xia, Liu Zhen1, Han Ming-lei, Hou Yong-lan 2016, 20 (23):  3445-3450.  doi: 10.3969/j.issn.2095-4344.2016.23.015 Abstract ( 342 )   PDF (1119KB) ( 250 )   Save BACKGROUND: Until now, it is still unclear whether the stem cell transplantation triggers adverse effects on the myocardial electrical activity, resulting in the emergence of arrhythmia. OBJECTIVE: To explore the effect of intracoronary transplantation of umbilical cord blood stem cells on arrhythmia score and incidence of ventricular arrhythmia. METHODS: According to therapeutic strategies, 73 patients with coronary heart disease were assigned to receive drug therapy in control group (n=38) and umbilical cord blood stem cell transplantation in observation group (n=35). Arrhythmia score, incidence of ventricular arrhythmia and adverse reactions were recorded and analyzed before and 1, 4, 8 weeks after transplantation. RESULTS AND CONCLUSION: After treatment, arrhythmia scores were significantly reduced in the two groups, especially in the observation group, to exhibit a continuous decline trend (P < 0.05). Compared with the control group, the incidence of ventricular arrhythmia was significantly lower in the observation (P < 0.05). However, there were no significant changes in the blood pressure, heart rate and blood oxygen saturation before and after transplantation, and no acute heart failure and death occurred in the two groups. These results suggest that the intracoronary transplantation of umbilical cord blood stem cells exhibits superiorities in the treatment of coronary artery disease, significantly reducing the arrhythmia score, reducing the incidence of ventricular arrhythmia, and resulting in less adverse reactions.   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 skin-derived mesenchymal stem cells from psoriasis lesions on T cell proliferation Wang Qing, Wang Da-hu, Wang Ai-xue, Guo Bing-shen, Wang Ning, Li Yu-ping 2016, 20 (23):  3451-3456.  doi: 10.3969/j.issn.2095-4344.2016.23.016 Abstract ( 450 )   PDF (1119KB) ( 264 )   Save BACKGROUND: Studies have found abnormalities in the biological activity and cytokine levels of bone marrow mesenchymal stem cells from psoriasis patients, while the biological activity of skin-derived mesenchymal stem cells from psoriasis lesions is rarely reported.  OBJECTIVE: To explore the influences of skin-derived mesenchymal stem cells from psoriasis lesions on T cell proliferation METHODS: After isolation and culture, skin-derived mesenchymal stem cells from psoriasis lesions and normal controls were separately co-cultured with peripheral blood T cells. T cell proliferation was detected using MTT assay, and levels of epidermal growth factor and transforming growth factor β1 in cell supernatant determined using ELISA method. RESULTS AND CONCLUSION: Skin-derived mesenchymal stem cells from both psoriasis lesions and normal controls significantly inhibited T cell proliferation (P < 0.05). Furthermore, skin-derived mesenchymal stem cells from psoriasis lesions exhibited a weaker inhibitory effect on T cell proliferation than normal skin-derived mesenchymal stem cells (P < 0.05). Skin-derived mesenchymal stem cells from psoriasis lesions significantly increased epidermal growth factor but reduced transforming growth factor β1 compared with the normal skin-derived mesenchymal stem cells (P < 0.05). In conclusion, maybe the imbalances of growth factor levels due to skin injury elicit a decrease in the inhibitory effect of skin-derived mesenchymal stem cells from psoriasis lesions on T lymphocytes.   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: isolation, identification and transplantation combined with chemotherapy for lung cancer in mice Zheng Tian-liang, Zhao Song, Guo Hai-zhou, Cui Guang-hui, Lin Da-wei 2016, 20 (23):  3457-3463.  doi: 10.3969/j.issn.2095-4344.2016.23.017 Abstract ( 342 )   PDF (4196KB) ( 241 )   Save BACKGROUND: Mesenchymal stem cells have pluripotent differentiation, and can promote cell engraftment and immune regulation. Therefore, we attempt to use human umbilical cord mesenchymal stem cells as a new source for treatment of lung cancer by exploring cell isolation, identification and transplantation combined with chemotherapy for lung cancer in mice. OBJECTIVE: To investigate the isolation and identification of human umbilical cord mesenchymal stem cells and its transplantation combined with chemotherapy for lung cancer in mice. METHODS: Human umbilical cord mesenchymal stem cells were isolated from fresh umbilical cord of newborns and identified using tissue culture and enzyme digestion. Twenty Balb/C nude mouse models of lung cancer were randomly divided into two groups: mice in chemotherapy group were given chemotherapy, and those in combined group given combination of chemotherapy with human umbilical cord mesenchymal stem cell transplantation. RESULTS AND CONCLUSION: Compared with the chemotherapy group, the gastrointestinal tract was rosy and shiny, intestinal mucosa was smooth and complete, and tumor mass and blood indexes significantly decreased in the combined group (P < 0.05). To conclude, mature human umbilical cord mesenchymal stem cells can be obtained by tissue culture and enzyme digestion, and the cell transplantation combined with chemotherapy can significantly reduce gastrointestinal tract damage and the make peripheral hemogram in a stable level.    中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Overexpression of microRNA-32 weakens the inhibition of polychlorinated biphenyls to the differentiation of P19 cells into cardiomyocytes Zhang Song-yu, Li Yan, Liu Jiang-bo, Li Gang 2016, 20 (23):  3464-3469.  doi: 10.3969/j.issn.2095-4344.2016.23.018 Abstract ( 237 )   PDF (3600KB) ( 220 )   Save BACKGROUND: Polychlorinated biphenyls inhibit the differentiation of P19 cells into cardiomyocytes. In the meanwhile, microRNAs play an important role in regulating cell differentiation. OBJECTIVE: To explore the effect of microRNA-32(miR-32) on the polychlorinated biphenyls-inhibited differentiation of P19 cells into cardiomyocytes. METHODS: P19 cells were co-cultured with 2.5 μmol/L polychlorinated biphenyls and 1% dimethyl sulfoxide. Afterwards, α-actinin, desmin, cTnI and GATA4 were identified by western blot assay. Wddwxrof polychlorinated biphenyls on the expression of miR-32 was evaluated by real-time PCR. Mouse eukaryotic vector expressing miR-32 was constructed by gene recombination technology, and transfected into P19 cells followed by co-cultured with 2.5 μmol/L polychlorinated biphenyls and 1% dimethyl sulfoxide. Then, expressions of differentiation-related proteins were detected by western blot assay. RESULTS AND CONCLUSION: Polychlorinated biphenyls inhibited the differentiation of P19 cells into cardio myocytes and decreased the miR-32 expression. Cell lines overexpressing miR-32 was successfully established in mice. Furthermore, we found that overexpression of miR-32 weakens the inhibition of polychlorinated biphenyls to the differentiation of P19 cells into cardiomyocytes.   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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Subcutaneous injection of basic fibroblast growth factors promotes the proliferation of hippocampal neural stem cells in rats with vascular dementia Liu Sen, Dai Zhen-xia, Wu Chun-fang, Hu Jing, Cui Bei 2016, 20 (23):  3470-3475.  doi: 10.3969/j.issn.2095-4344.2016.23.019 Abstract ( 295 )   PDF (1067KB) ( 345 )   Save BACKGROUND: As a neurotrophic factor, basic fibroblast growth factors extensively distribute in the central nervous system, and play an important physiological role by combination with their relative receptors. OBJECTIVE: To investigate the effect of basic fibroblast growth factors on the learning ability and proliferation of nestin-positive hippocampal neural stem cells in rats with vascular dementia. METHODS: Totally 30 Sprague-Dawley rats were randomly divided into vascular dementia, sham operation and treatment groups. The vascular dementia and treatment groups were for preparing vascular dementia model, and the treatment group was given subcutaneous injection of basic fibroblast growth factors. Subsequently, at 4 weeks, the learning ability of rats and the number of nestin-positive hippocampal neural stem cells was detected by the Morris water maze test and immunohistochemical staining, respectively. RESULTS AND CONCLUSION: Compared with the vascular dementia group the latency period was significantly shorter in the sham operation and treatment groups, and the number of times crossing the target quadrant was significantly higher in the sham operation and treatment groups (P < 0.05). But there was no significant difference between the sham operation and treatment groups (P > 0.05). Under fluorescence microscope, yellow-green fluorescence stained neurons positive for basic fibroblast growth factor could be found in the CA1, CA2 and CA3 of the treatment group. Additionally, the number of nestin-positive neurons in the CA1 area of the hippocampus was the most in the treatment group, followed by the sham operation group, and the least in the vascular dementia group. These results suggest that the subcutaneous injection of basic fibroblast growth factors can migrate to the hippocampus, , and improve the learning ability of rats by inducing proliferation of nestin-positive hippocampal neural stem cells.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Basic fibroblast growth factors promote proliferation of endogenous neural stem cells after spinal cord injury in mice Li Yu-bo, Ding Li-xiang 2016, 20 (23):  3476-3483.  doi: 10.3969/j.issn.2095-4344.2016.23.020 Abstract ( 224 )   PDF (1299KB) ( 254 )   Save BACKGROUND: So far steroid pulse therapy and surgical decompression are the main accepted therapies for spinal cord injury, but these methods make no effects on injured neurons. Nerve growth factors and endogenous neural stem cells play a role in the neuronal regeneration and remyelination, which provides a new idea for spinal cord injury treatment. OBJECTIVE: To explore the effect of the basic fibroblast growth factor on proliferation of endogenous neural stem cells after spinal cord injury and to analyze its relationship with the increased fluoro-gold labeled neurons. METHODS: Totally 48 Kunming mice were randomly divided into four groups including normal, spinal cord injury, treatment and sham operation groups. Acute spinal cord injury models were established in the spinal cord injury and treatment groups, and the normal group was subjected to operation that did not damage the spinal cord. At 2 hours after regaining consciousness, the treatment group was given daily injection of MTPBS containing 25 μg/kg basic fibroblast growth factors and 1% album. And at 7 days, laminectomy was carried out again at the L1 segment in the former three groups and a small piece of sterile gelfoam soaked with fluoro-gold was inserted into the incision. The sham operation group was given no processing. Afterwards, mouse motor behavior was assessed using Rotarod and Platform Hang tests; neurons in the corticospinal and rubrospinal tracts were labeled with fluoro-gold; the number of endogenous neural stem cells positive for nestin was detected by immunohistochemistry method. Besides, the correlation between the number of fluoro-gold labeled neurons and the number of endogenous neural stem cells was assessed. RESULTS AND CONCLUSION: The basic fibroblast growth factor could significantly improve the mouse motor behavior after spinal cord injury. And the number of endogenous neural stem cells was significantly increased after the basic fibroblast growth factor injection, which was related to the increased fluoro-gold labeled neurons. In conclusion, basic fibroblast growth factors play an important role in the proliferation of endogenous neural stem cells after spinal cord injury. Furthermore, endogenous neural stem cells improve locomotive behaviors by encouraging the neuronal proliferation.    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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Insight into stem cells, microenvironment and methods for promoting blood circulation and removing stasis  Zhang Jin-sheng, Zhang Bao-xia, Zhu Hui-fang, Zhang Yang-yang 2016, 20 (23):  3484-3490.  doi: 10.3969/j.issn.2095-4344.2016.23.021 Abstract ( 279 )   PDF (1126KB) ( 239 )   Save BACKGROUND: There is a certain association between stem cells, microenvironment and methods for promoting blood circulation and removing stasis. OBJECTIVE: To propose a perspective of “kidney properties” of promoting blood circulation and removing stasis by elaborating the correlation and interactions of stem cells and microenvironment with methods for promoting blood circulation and removing stasis. METHODS: A computer-based search of Chinese Journal Full-text Database, Wanfang and PubMed was performed for articles published form 2009 to 2015 addressing improving stem cell proliferation and differentiation by promoting blood circulation and removing stasis as well as microenvironments. The keywords were “bone mesenchymal stem cells, transformation, differentiation, blood-activation and stasis-resolved method, microenvironment” in Chinese and English, respectively. Finally, 32 literatures were reviewed. RESULTS AND CONCLUSION: Stem cells homing to the damaged tissue trigger an “environment- dependent differentiation”, that is, stem cells can differentiate into myocardial cells, vascular endothelial cells and other injured tissues and cells under local special microenvironments. Microenvironment as an assembly for integrating dynamic, microscopic changes as well as development of a variety of indices is of great importance for stem cell survival, proliferation and differentiation. Recipes for promoting blood circulation and removing stasis are involved to regulate multi-steps and multi-targets during stem cell homing. Theoretically, this review attempts to investigate correlation among stem cells, microenvironment and methods for promoting blood circulation and removing stasis in association with previous preclinical and clinical findings. Here, we further refine the view of “kidney properties” of promoting blood circulation and removing stasis and clarify that stem cells as the material basis of “kidney properties” of promoting blood circulation and removing stasis is the main executor for promoting blood circulation and removing stasis. This paper is of a certain value for fully revealing the scientific connotation of the theory of promoting blood circulation and removing stasis.    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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Stem cells in the skin regeneration and wound healing: problem and perspective Fang Shao-xia, Guo Fen 2016, 20 (23):  3491-3496.  doi: 10.3969/j.issn.2095-4344.2016.23.022 Abstract ( 347 )   PDF (1014KB) ( 267 )   Save BACKGROUND: Present treatments for chronic skin wounds have certain limitations, and adult stem cells play a potential part in cutaneous repair and regeneration. OBJECTIVE: To review effects of stem cells in skin regeneration and wound healing. METHODS: The first author retrieved CNKI and Medline databases by computer for relevant articles published from 2000 to 2010. The keywords were “epidermal stem cells, hair follicle stem cells, stem cells, transplantation, dermal stem cells” in Chinese and in English, respectively. Then totally 489 papers were obtained after initial survey, and according to the inclusion criteria, 30 articles were selected for review. RESULTS AND CONCLUSION: Epidermal stem cells and other adult stem cells have been applied to treat wounds and other skin diseases. Epidermal stem cells are the crucial cell source of skin development, repair and remodeling. Epidermal stem cells are always in a resting state in vivo. Unless, skin injure or culture in vitro, cell division and proliferation will be significantly fastened. The stability of the epidermis mainly depends on the asymmetric division of a subpopulation, in which two daughter cells are produced, including one with characteristics of stem cells, and the other differentiated into transient amplifying cells that will be differentiated into post mitotic cells after a series of cell divisions (3-5 times). Afterwards, those post mitotic cells are developed into terminal differentiation cells on the basal layer, finally detach from the epidermis as dander. In addition, it is unclear whether epidermal factors are related to apoptosis, migration and differentiation in the process of wound repair and even under physiological conditions. Therefore, application of stem cells in wound healing requires a further discussion.    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