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05 November 2014, Volume 18 Issue 45 Previous Issue    Next Issue

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Role of Toll-like receptor 3 in the immunoregulatory function of embryonic bone marrow mesenchymal stem cells on Th17 cells Guo Zhen-xing, Chen Zhen-ping 2014, 18 (45):  7217-7221.  doi: 10.3969/j.issn.2095-4344.2014.45.001 Abstract ( 284 )   PDF (421KB) ( 488 )   Save BACKGROUND: Previous studies have found that embryonic bone marrow mesenchymal stem cells can promote human Th17 cell proliferation, but the inherent regulatory mechanisms still need to be elucidated.   OBJECTIVE: To investigate the role of Toll-like receptor 3 in the immunoregulation of Th17 cells by mesenchymal stem cells. METHODS: Human CD4+ T cells from healthy donors were isolated by immunomagnetic bead method, and then cultured alone or co-cultured with embryonic bone marrow mesenchymal stem cells for 4 days. The mRNA expression level of interleukin-17, Toll-like receptor 3 and MyD88 was detected by real-time PCR. RESULTS AND CONCLUSION: Compared with CD4+ T cell cultured alone group, the mRNA level of interleukin-17 was significantly higher in the co-culture group (3.59±0.11 vs. 1.14± 0.08, P < 0.01). Consistent with the expression of interleukin-17 mRNA, increased level of Toll-like receptor 3 mRNA was detected in the co-culture group compared with the CD4+ T cell cultured alone group (3.10±1.60 vs. 0.94± 0.01, P < 0.05). Furthermore, MyD88 in the co-culture group was significantly higher than that in CD4+ T cell cultured alone group (2.29±0.05 vs. 1.85±0.31, P < 0.01). Toll-like receptor 3 may be involved in the immunoregulation of Th17 cells by embryonic bone marrow mesenchymal stem cells, which provides experimental evidence for potential cell therapeutic strategy. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Expansion of chicken bone marrow mesenchymal stem cells by laminin culture system Li Shuang-xing, Qi Yuan, Piao Feng-yuan, Li Ya-chen, Liu Xiao-hui, Shao Jing, Li Shuang-yue 2014, 18 (45):  7222-7226.  doi: 10.3969/j.issn.2095-4344.2014.45.002 Abstract ( 295 )   PDF (753KB) ( 561 )   Save BACKGROUND: Bone marrow mesenchymal stem cells from chickens are important cell models for embryonic developmental biology, immunology and oncology research. However, it is difficult to keep bone marrow mesenchymal stem cells with good undifferentiated potential in a large-scale expansion system. OBJECTIVE: To establish a culture system in vitro with laminin coating to expand bone marrow mesenchymal stem cells from chickens. METHODS: Isolated bone marrow mesenchymal stem cells from chickens were seeded in laminin-coated plates and traditional two-dimensional plates, respectively. After expansion in vitro, the morphological characteristics, expression of surface markers, expansion characteristics and adipogenic differentiation of bone marrow mesenchymal stem cells in both conditions were analyzed and compared. RESULTS AND CONCLUSION: There were no statistical differences in the morphological characteristics and expression of surface markers of bone marrow mesenchymal stem cells expanded by laminin-coated plates and traditional two-dimensional plates. But, the expansion characteristics and adipogenic differentiation of bone marrow mesenchymal stem cells cultured in laminin-coated plates were better than those in traditional two-dimensional plates. Laminin culture system could quickly amplify out of a large number of chicken bone marrow mesenchymal stem cells with better proliferation ability and undifferentiated performance. All above results indicated that a more efficient expansion system with laminin coating is established. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effects of three-dimensional spheroid culture system on biological characteristics of mouse bone marrow mesenchymal stem cells He Xin, Li Xue, Bao Hui-jing, Wang Ren-feng, Liu Yun-de, Song Shi-wei 2014, 18 (45):  7227-7232.  doi: 10.3969/j.issn.2095-4344.2014.45.003 Abstract ( 346 )   PDF (539KB) ( 529 )   Save BACKGROUND: Bone marrow mesenchymal stem cells have attracted widespread attention for the capabilities of self-renewal and muti-differentiation, which have been used in treatment of various diseases.   OBJECTIVE: To study the effect of three-dimensional spheroid culture system on the stemness and senescence of bone marrow mesenchymal stem cells.   METHODS: Mesechyaml stem cells were isolated from the bone marrow of C57/B6 mice, 3 weeks old, and cultured onto the culture plates coated with or without chitosan. After 5 days of culture, the cell phenotype and expression of stemness related markers CD44 and Sca-1 were analyzed by flow cytometry. PI and Annexin-V staining were used to detect cell apoptosis. Also, β-Gal staining was applied for identification of aging.   RESULTS AND CONCLUSION: The mouse mesenchymal stem cells began to form spheroids on day 3. The stemness-related markers, including CD44 and Sca-1, expressed higher in spheroid mesenchymal stem cells than the cells under normal culturing. Compared with the normal culture group, the apoptosis and senescence of cells from spheroid culture were lower. The results indicate that the formation of spheroids on chitosan films can increase the stemness, decrease the apoptosis and slow the senescence of mesenchymal stem cells.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effect of platelet-rich fibrin released transforming growth factor beta 1 on the proliferation of bone marrow mesenchymal stem cells in vitro Chen Cheng, Li Shu-hui, Zhang Wen-li, Li Yi-ming, Zhou Jing, Wu Pei-ling 2014, 18 (45):  7233-7238.  doi: 10.3969/j.issn.2095-4344.2014.45.004 Abstract ( 298 )   PDF (410KB) ( 558 )   Save BACKGROUND: Bone marrow mesenchymal stem cells are the ideal cells for tissue repair. Whether the ability of in vitro proliferation can be enhanced is a key factor to promote tissue repair. OBJECTIVE: To observe the effect of transforming growth factor β1 on the proliferation of bone marrow mesenchymal stem cells in vitro. METHODS: Blood samples were taken from the central artery of rabbits to prepare platelet-rich fibrin by centrifugation method which was then placed into fresh DMEM at 37 ℃ for 7, 14, 21, 28 days to collect exudates. The mass concentrations of transforming growth factor-β1 in the exudates of platelet-rich fibrin were detected. Rabbit bone marrow mesenchymal stem cells were collected and cultured in the conditioned medium made by the exudates of platelet-rich fibrin, and the proliferation of cells was observed. RESULTS AND CONCLUSION: Concentration of transforming growth factor β1 was increased with time increasing, increased fastest at 21-28 days, and peaked at 28 days. Under the same stimulus concentration, the proliferation of bone marrow mesenchymal stem cells was reduced at 0-1 day, increased obviously at 1-2 days, and entered into a steady phase at 2-3 days. Under 150 ng/L transforming growth factor β1, bone marrow  mesenchymal stem cells proliferated fastest. Experimental findings indicate that with the increase of time, the concentration of transforming growth factor β1 in the exudates of platelet-rich fibrin increase gradually, and the conditioned media containing different concentrations of transforming growth factor β1 play different roles in promoting the proliferation of bone marrow mesenchymal stem cells. Bone marrow mesenchymal stem cells cultured in the conditioned medium containing 150 ng/L transforming growth factor β1 for 2-3 days can proliferate fastest. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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The anti-aging study of human umbilical cord mesenchymal stem cells combined with lycopene treatment in the aging beagles Wang Lin-lin, Cui Xiao-lan, Shi Han, Cheng Xue, Liu Jia, Shen Yi, Li Qian-qian, Wang Yi-zhong 2014, 18 (45):  7239-7245.  doi: 10.3969/j.issn.2095-4344.2014.45.005 Abstract ( 354 )   PDF (743KB) ( 633 )   Save BACKGROUND: Previous tudies have shown that the anti-aging effects of stem cells with lycopene are more significant, and can also significantly improve the aging body immune function. OBJECTIVE: To investigate the anti-aging effects of human umbilical cord mesenchymal stem cells combined with lycopene on the aging beagles. METHODS: Sixteen aging beagles (6-7 years old) were randomly divided into two groups: aging control group and aging treatment group; young beagles (3-4 years old) were chosen as young control group. In the aging treatment group, human umbilical cord mesenchymal stem cells combined with lycopene was given; while in the other two group, an equal amount of DMEM/F12 cell culture medium and sunflower oil was given. Each dog's general conditions were observed regularly during the whole progress. The changes of superoxide dismutase, malondialdehyde, glutathione peroxidase in the serum were detected at regular time of the whole process, and the structure changes of each organ were observed at 24 weeks of treatment. RESULTS AND CONCLUSION: (1) Before treatment, the levels of superoxide dismutase and glutathione peroxidase in the aging control and treatment groups were lower than those in the young control group (P < 0.05), while the malondialdehyde content was higher than that in the young control group (P < 0.05). However, there was no significant statistical difference between aging control and treatment groups (P > 0.05). (2) For the aging treatment group at 24 weeks of treatment: the beagle fur became clearer and smoother, motility was strengthened, appetite became better; and the activity of superoxide dismutase in serum at 8 to 24 weeks of treatment increased significantly compared with before treatment (P < 0.05), the activity of glutathione peroxidase significantly increased from 6 weeks (compared with treatment before, P < 0.05), the malondialdehyde content decreased significantly from 4 weeks of treatment to the completion of the experiment (compared with before treatment, P < 0.05). (3) After the experiment, the microscopic observation showed that compared with the aging control group, the tissues and organ structures of the aging treatment group were all clear, had no inflammatory infiltrates, no obvious necrosis and fibrosis lesions. These results were mainly consistent with the observations of young control group. The above results show that umbilical cord mesenchymal stem cells combined with lycopene therapy on the natural aging beagles may enhance the activities of antioxidant enzymes, reduce malondialdehyde content, and their combination also can repair tissue structures and promote the functions, which has obvious anti-aging effects. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Optimized medium accelerates differentiation of bone marrow mesenchymal stem cells Li Qian, Yang Jun 2014, 18 (45):  7246-7249.  doi: 10.3969/j.issn.2095-4344.2014.45.006 Abstract ( 304 )   PDF (319KB) ( 557 )   Save BACKGROUND: Studies have found that bone marrow mesenchymal stem cells, under certain conditions, can be induced to differentiate into neurons and glial cells, which to some extent solves the problem of the source of seed cells. Induction methods currently used are different, and their efficiencies are not the same.   OBJECTIVE: To observe the effects of different antioxidants on differentiation of rat bone marrow mesenchymal stem cells into neuron-like cells in vitro. METHODS: Bone marrow mesenchymal stem cells from Wistar rats were divided into four groups: non-intervention group, β-mercaptoethanol group, retinoic acid group, β-mercaptoethanol+retinoic acid group. Changes in cell morphology and positive rate of neuron-specific enolase and microtubule-associated protein 2 were observed and detected at 5 hours, 12 hours, 1 day, 3 days, 5 days, 7 days, and 10 days after induction. RESULTS AND CONCLUSION: Except non-intervention group, bone marrow mesenchymal stem cells in the other three groups were gradually becoming spindle-shaped, and gave birth to many small protrusions that were interconnected into a network, showing neuron-like cell morphology. Immunocytochemical staining showed that the efficiency of the β-mercaptoethanol+retinoic acid group was the highest at 10 days after induction, and the positive rates of neuron-specific enolase and microtubule-associated protein 2 were 71.63% and 79.72%, respectively. The results show that β-mercaptoethanol can be combined with retinoic acid to accelerate the differentiation of bone marrow mesenchymal stem cells into neuron-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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Rat’s Schwann cells induce neural differentiation of human umbilical cord mesenchymal stem cells Zhu Jia-xue, Tao Hai-rong, Shen Zun-li 2014, 18 (45):  7250-7254.  doi: 10.3969/j.issn.2095-4344.2014.45.007 Abstract ( 243 )   PDF (688KB) ( 741 )   Save BACKGROUND: Mesenchymal stem cells can differentiate into nerve cells by chemical induction or co-culture method, but whether mesenchymal stem cells co-cultured with Schwann cells differentiate into neuronal-like cells or Schwann-like cells is still controversial. OBJECTIVE: To explore the inductive role of Schwann cells derived from rats in the differentiation of human umbilical cord mesenchymal stem cells. METHODS: Cocultures of human umbilical cord mesenchymal stem cells (1×109/L) and Schwann cells (1×109/L) from neonatal rats were performed using transwell culture dishes. After 2 weeks of cocultures, morphology of the cultured human umbilical cord mesenchymal stem cells was observed, and the phenotypic changes of cells were also detected with immumocytochemistry techniques. RESULTS AND CONCLUSION: After 2 weeks of cocultures, some differentiated cells showed neuron-like morphology, and expressed nestin, NF-200 and β-III-tubulin, but did not express Schwann cell special marker S100 and oligodendrocytes special marker MAB1580. These findings indicate that human umbilical cord mesenchymal stem cells can transdifferentiate into neuronal-like cells by cocultures with rat’s Schwann cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Differentiation of embryonic stem cells into endothelial progenitor cells under hypoxic condition Xia Zhen-wei, Wang Ji-wen, Li Xiang-dong, Wei Guo-feng 2014, 18 (45):  7255-7259.  doi: 10.3969/j.issn.2095-4344.2014.45.008 Abstract ( 259 )   PDF (410KB) ( 610 )   Save BACKGROUND: The quantity and quality of seed cells is a critical bottleneck of the development of vascular tissue engineering. To address this issue, stem cell-derived endothelial cells have been a hot spot in this field due to their potential in providing the ideal seed cells. OBJECTIVE: To elucidate the effect of vascular endothelial growth factor (VEGF) supplementation combined with hypoxic culture condition on the lineage-specific differentiation of embryonic stem cells into endothelial cells. METHODS: Serum-free medium mTeSR®1 was applied to cultivate H9 cells in vitro. A conditioned medium containing 50 μg/L vascular endothelial growth factor was utilized to induce H9 cells to differentiate into endothelial cells under the hypoxic culture condition (5% O2). The cell under normal condition (5% CO2) with or without vascular endothelial growth factor served as controls. The phenotype and function of human embryonic stem cells-derived endothelial cells were assayed by immunofluorescence staining, quantitative RT-PCR, and low-density lipoprotein uptake experiment. RESULTS AND CONCLUSION: Compared with the control group, the H9 cells were induced to be differentiated into endothelial-like cells more efficiently when they were cultivated under a conditioned medium with vascular endothelial growth factor supplementation under the hypoxic condition. These differentiated cells not only expressed some important surface markers of endothelia cells, including kdr, pecam, but also took in low-density lipoprotein to form microvessle-like structures. This culture system supports a synergy effect of vascular endothelial growth factor and hypoxic environment that can efficiently promotes the lineage-specific differentiation of embryonic stem cells into endothelial cells with good phenotype and functionality. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Culture and identification of SK-N-SH neuroblastoma stem cells Xie Yan, Su Yu-xi, Qin Jia-qiang, Nan Guo-xin, Huang Shi-feng, Wang Zhong-liang, Cai Wen-quan 2014, 18 (45):  7260-7265.  doi: 10.3969/j.issn.2095-4344.2014.45.009 Abstract ( 438 )   PDF (1134KB) ( 733 )   Save BACKGROUND: Neuroblastoma is a common solid tumor in children. Pediatric tumors are little affected by environmental factors, but closely related to child development. The suspension method is an effective and reliable method to harvest neoplastic stem cells. OBJECTIVE: To culture the cell clones of human neuroblastoma cell line SK-N-SH and to assess the biological properties of the cell clones. METHODS: Using the suspension method with no serum media, tumor cell clones were obtained. Immunofluorescence method was used to identify whether tumor cell clones exhibit stem cell properties. SK-N-SH neuroblastoma was labeled by luciferase, and tumor cell clones and tumor cells were seeded onto the back of nude mice to monitor cell proliferative properties in nude mice using in vivo imaging. RESULTS AND CONCLUSION: Using the suspension culture method, SK-N-SH neuroblastoma cells could successfully develop into cloning balls. Under serum-free culture, cloning balls were immunofluorescently used to detect molecular markers that showed strong positive expression. Cloning balls subcutaneously implanted into nude mice showed the strong ability of self-renewal and differentiation as stem cells. The cell clones cultured by the suspension method strongly expressed Nestin, but weakly expressed glial fibrillary acidic protein, neuron-specific tubulin, possessing stem cell characteristics and strong proliferation and metastasis in nude mice. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Isolation and identification of neural stem cells from newborn mouse hippocampus, olfactory bulb and cortex Ma Jun-ning, Gao Jun-wei, Hou Bo-ru, Ren Hai-jun, Chen Si-hua, Liu Ji-xing, Yan Gui-zhong 2014, 18 (45):  7266-7272.  doi: 10.3969/j.issn.2095-4344.2014.45.010 Abstract ( 392 )   PDF (590KB) ( 679 )   Save BACKGROUND: To in vitro isolate neural stem cells with high purity and uniform biological properties and to establish a complete set of neural stem cell culture system is the basis for neural stem cell research. OBJECTIVE: To establish an isolation and culture system for neural stem cells from newborn mouse hippocampus, olfactory bulb and cortex and to analyze the biological properties of cells. METHODS: Neural stem cells were isolated from the hippocampus, olfactory bulb and cortex tissue of newborn Kunming mice by mechanical separation and trypsin digestion. Serum-free culture technology, mechanical pipetting and trypsin digestion were used for subculture of neural stem cells. 10% fetal bovine serum was used to induce differentiation of neural stem cells. Neural stem cells and their differentiated products were identified by immunofluorescent staining of Nestin, CD133, β-TubulinIII, glial fibrillary acidic protein. RESULTS AND CONCLUSION: The neural stem cell obtained from newborn mouse hippocampus, olfactory bulb and cortex had the capacity of self-renewal and differentiation which were positive for Nestin and CD133. After induction with fetal bovine serum, neural stem cell could differentiation to β-tubulinIII or glial fibrillary acidic protein positive cells that were neurons and astrocytes. This experiment has successfully established the neural stem cell isolation, culture, identification and induction system, providing experimental basis for subsequent studies of neural stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Association between some cytokines and graft-versus-host disease after allogeneic hematopoietic stem cell transplantation for beta-thalassemia major Chen Li-bai, Wen Jian-yun, Ruan Yong-sheng, Pei Fu-yu, Liu Hua-ying, He Yue-lin, Li Chun-fu, 2014, 18 (45):  7273-7278.  doi: 10.3969/j.issn.2095-4344.2014.45.011 Abstract ( 327 )   PDF (362KB) ( 530 )   Save BACKGROUND: Cytokines play an important role in the occurrence and development of graft-versus-host disease, but there is a current lack of reports on the association between cytokines and graft-versus-host disease after allogeneic hematopoietic stem cell transplantation for treatment of β-thalassemia major. OBJECTIVE: To investigate the association between cytokines and graft-versus-host disease after allogeneic hematopoietic stem cell transplantation for β-thalassemia major. METHODS: We observed the dynamic variation of interleukin 6, interleukin 8, interleukin 12, tumor necrosis factor-α and macrophage migration inhibitory factor in 11 children with β-thalassemia major before onset of graft-versus-host disease, when graft-versus-host disease occurred, at days 4 and 7 after onset of graft-versus-host disease, and when graft-versus-host disease disappeared. RESULTS AND CONCLUSION: There was a significant difference in serum levels of interleukin-6, interleukin-12, tumor necrosis factor-α, macrophage migration inhibitory factor in different time points, and the highest levels of different cytokines appeared when graft-versus-host disease occurred, followed by those at 7 days after  graft-versus-host disease. There was a significant difference in serum levels of interleukin-8 in different time points, and the highest level appeared at 4 days after graft-versus-host disease. The dynamic expression of interleukin-6, interleukin-8, interleukin-12, tumor necrosis factor-α, macrophage migration inhibitory factor can estimate the immune function of β-thalassemia major patients who develops graft-versus-host disease after allogeneic hematopoietic stem cell transplantation, and can be used as the immunobiology indicators for the early diagnosis of graft-versus-host disease. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Variation of Th1/Th2 and Terg in rheumatoid arthritis patients undergoing umbilical cord mesenchymal stem cell transplantation Wang Li-ming, Wang Li-hua, Li Ming, Bai Wen, Zhong Zhan-qiang, Shi Jun, Zhou Jian-jun, 2014, 18 (45):  7279-7284.  doi: 10.3969/j.issn.2095-4344.2014.45.012 Abstract ( 411 )   PDF (333KB) ( 718 )   Save BACKGROUND: Rheumatoid arthritis is an autoimmune disease, and traditional treatment methods are difficult to effectively solve the patient's lack of immune tolerance mechanisms. With the development of stem cells in regenerative medicine, stem cell therapy has become a hot spot in the treatment of autoimmune diseases.   Currently, studies on cell transplantation for the treatment of rheumatoid arthritis are rarely reported. OBJECTIVE: To study the influence of umbilical cord mesenchymal stem cell therapy on the changes of Th1/Th2 and Treg in rheumatoid arthritis patients, thereby seeking new therapies for rheumatoid arthritis. METHODS: We selected 180 cases of rheumatoid arthritis, including 27 patients as control group undergoing non-steroidal anti-inflammatory drugs and anti-rheumatic drugs and 153 patients as cell treatment group undergoing intravenous infusion of 40 mL umbilical cord mesenchymal stem cells at a density of 4×107. Dosing regimen was same in the two groups. The 76 of 153 patients accepted second cell therapy at 3-4 months after the first cell therapy. After follow-up of 3 and 6 months, clinical effectiveness evaluation (DAS28, HAQ, ACR20), rheumatoid factor, anti-CCP antibodies, T cell subsets, Th cytokine were detected; for patients with second cell therapy, T cell subsets and Treg were detected at 8 months after treatment. RESULTS AND CONCLUSION: (1) At 3 months after treatment, the DAS28, HAQ and ACR20 scores were significantly lower in the cell treatment group than the control group (P < 0.01). (2) At 3 and 6 months after cell therapy, the DAS28 and HAQ scores were significantly decreased in the cell treatment group (P < 0.01), and these scores were decreased continuously after second cell therapy (P < 0.01). (3) Interferon-γ level in cells did not change obviously at 3-6 months after treatment, but the interleukin-4 level was gradually increased at 6 months after treatment (P < 0.05). (4) The number of Treg cells was significantly increased at 3-6 months after treatment (P < 0.01), which was closely related to ACR, especially ACR70 percentage (P < 0.05); the ratio of CD4+ Treg was increased significantly at 3 months after treatment (P < 0.05), and this increasing trend was also maintained at 6 and 8 months after treatment, but there was no significant difference (P > 0.05). (5) B cell levels were significantly decreased at 6 months after treatment (P > 0.05); the rheumatoid factor value was significantly decreased at 3-6 months after treatment (P < 0.05). (6) There was no change in anti-CCP antibody and interleukin-17 levels at 3-6 months after treatment. These findings indicate that after treatment with umbilical cord mesenchymal stem cells, the Th1/Th2 tends to balance and Treg level is elevated in rheumatoid arthritis patients, which are directly related to clinical trials and symptomatic relief. Therefore, standard rheumatism medication combined with umbilical cord mesenchymal stem cell transplantation can improve immune network effects, adjust the immune tolerance and prevent illness progress in rheumatoid arthritis patients. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | Related Articles | Metrics

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Dopamine content in the stratum of Parkinson’s disease rats after transplantation of tyrosine hydroxylase-modified human umbilical cord blood mesenchymal stem cells Fan Zhi-gang, Qiao Lei 2014, 18 (45):  7285-7289.  doi: 10.3969/j.issn.2095-4344.2014.45.013 Abstract ( 261 )   PDF (812KB) ( 662 )   Save BACKGROUND: Stem cell therapy is superior to drug therapy for recovery of patient’s physiological mode, and cell transplantation therapy is becoming a trend.   OBJECTIVE: To observe the changes in dopamine content in the stratum of Parkinson’s disease rats after transplantation of tyrosine hydroxylase-modified human umbilical cord blood mesenchymal stem cells. METHODS: After identification by enzyme digestion, pEGFP-C2-TH plasmid was transfected into the fourth generation of human umbilical cord blood mesenchymal stem cells by electroporation method, and then transfected cells were injected into the right cerebral ventricle of Parkinson’s disease rats (experimental group). PBS injection was performed in the control group. Migration of dopamine in the brain tissue of rats was observed, and the content of dopamine was detected by high performance liquid chromatography． RESULTS AND CONCLUSION: At 8 weeks after cell transplantation, the cells gradually migrated to the ventricles; after 12 weeks, the cells migrated to the cortex, and expressed tyrosine hydroxylase antigen. Meanwhile, the content of dopamine was significantly higher in the experimental group than the control group (P < 0.05). These results reveal that the intraventricular transplantation of tyrosine hydroxylase-modified human umbilical cord blood mesenchymal stem cells has obvious therapeutic effect on Parkinson’s disease rats. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effect of acupoint injection of bone marrow mesenchymal stem cells on expression of vascular cell adhesion molecule-1 and very late antigen-4 in myocardial infarction model rats   Chen Yan, Yang Guan-lin, Bai Xue-song, Zhang Zhe, Guan Xue-feng, Sui Ji-feng 2014, 18 (45):  7290-7293.  doi: 10.3969/j.issn.2095-4344.2014.45.014 Abstract ( 381 )   PDF (420KB) ( 491 )   Save BACKGROUND: Now, the bone marrow mesenchymal stem cell homing is thought to be mediated by adhesion molecules and chemokines, and this process involves bone marrow endothelial cells, hematopoietic stem cells, bone marrow microenvironment and its secreted or expressed molecules, in which, adhesion molecules may play an important role. OBJECTIVE: To explore the migrating and chemotactic mechanism of bone marrow mesenchymal stem cells via acupoint injection into myocardial cells by determining the expression of vascular cell adhesion molecule-1 and very late antigen-4. METHODS: Bone marrow mesenchymal stem cells were cultured using adherent method, and the passage 3 cells were used as seed cells at a density of 1×1010/L. Sixty Sprague-Dawley rats were randomly divided into sham group, model group, myocardial injection group, and acupoint injection group, 15 rats in each group. The left coronary arteries of rats were ligated for establishing a model of myocardial infarction. At 72 hours after myocardial infarction, 0.3 mL bone mesenchymal stem cells were transplanted into the Xinyu, Zhiyang, Tanzhong acupoints, respectively, in the acupoint injection group; while in the myocardial injection group, secondary thoracotomy was done, and 1.2 mL bone marrow mesenchymal stem cells were equably transplanted into six sites in the feeding area of the left anterior descending artery and the surrounding myocardium. At 4 weeks after myocardial infarction, a multi-channel polygraph was adopted for detection of hemodynamic parameters, and the levels of serum vascular cell adhesion molecule-1 and very late antigen-4 were measured by ELISA. RESULTS AND CONCLUSION: The heart function of rats in the myocardial injection and acupoint injection groups were improved, and compared with the model group, the levels of serum vascular cell adhesion molecule-1 and very late antigen-4 were significantly higher in the myocardial injection and acupoint injection groups. However, there was no significant difference between the myocardial injection and acupoint injection groups. These findings suggest that vascular cell adhesion molecule-1 and very late antigen-4 may be one of the chemotactic mechanisms of bone mesenchymal stem cells transplanted in myocardial infarction model rats by acupoint injection. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Mechanism of ultrasound-mediated microbubbles enhancing homing effect of transplanted mesenchymal stem cells Qian Jian, Chen Fei, Fan Guo-feng, Sha Du-juan, Wang Lu-na, Li Qi-ming, Ma Hao, Chen Yi-bing, Zhang Jun 2014, 18 (45):  7294-7298.  doi: 10.3969/j.issn.2095-4344.2014.45.015 Abstract ( 267 )   PDF (330KB) ( 502 )   Save BACKGROUND: Animal studies have indicated ultrasound-mediated microbubbles can significantly enhance the effect of stem cell transplantation to treat ischemic diseases. But its mechanism is still unknown. OBJECTIVE: To explore the mechanism of ultrasound-mediated microbubbles to significantly enhance the effect of stem cell transplantation in the treatment of ischemic diseases. METHODS: Bone marrow mesenchymal stem cells and vascular endothelial cells of rats were cultured in vitro, and then randomized to three groups: control group with no intervention, ultrasound group exposed to ultrasound at 1 MHz, 1 W/cm2 for 90 seconds, and ultrasound-mediated microbubble group treated with 5 μL liposomes ultrasound microbubbles containing fluorocarbon gases (about 2×1011/L) and ultrasound exposure at 1 MHz, 1 W/cm2 for 90 seconds. RESULTS AND CONCLUSION: Compared to the control group, ultrasound-mediated microbubbles significantly increased expressions of vascular endothelial growth factor and stromal cell-derived factor 1 in the supernatant of vascular endothelial cells (P < 0.05); ultrasound had no effect on the expression of vascular endothelial growth factor, but decreased the level of stromal cell-derived factor 1 (P < 0.01). Ultrasound-mediated microbubbles and the ultrasound alone could significantly enhance the CXCR4 gene expression in bone marrow mesenchymal stem cells as compared with the control group (P < 0.01), but there was no difference between the ultrasound-mediated microbubble group and the ultrasound group (P > 0.05). These findings suggest that 1 W/cm2 ultrasound-mediated microbubbles can promote vascular endothelial growth factor and stromal cell-derived factor 1 secretion by vascular endothelia cells, and meanwhile promote CXCR4 gene expression in bone marrow mesenchymal stem cells. This may be the mechanism of the ultrasound-mediated microbubbles enhancing homing effect of transplanted stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Transfection of stem cells derived from rat dental pulp with green fluorescent protein infection by lentiviral vector Zhang Rui-han, Liu Jia, Nie Shan-shan, Wang Xuan, Li Bo-qi, Sun Da-lei, Refukati Dilimaolati, Liu Yi-shan 2014, 18 (45):  7299-7305.  doi: 10.3969/j.issn.2095-4344.2014.45.016 Abstract ( 291 )   PDF (1899KB) ( 457 )   Save BACKGROUND: Stable and efficient labeling of dental pulp stem cells in vitro is most important in tracer technique, which is also the basis of tooth regeneration in vivo. OBJECTIVE: To determine the optimal condition and method for transfection of stem cells derived from rat dental pulp with green fluorescent protein infection by lentiviral vector and to determine whether green fluorescent protein-labeled dental pulp stem cells maintain their stem cell properties. METHODS: Rat dental pulp stem cells were obtained by modified enzyme digestion method, to identify the immune phenotype and differentiation potential. Dental pulp stem cells were infected with green fluorescent protein by lentiviral vector for 24 and 48 hours at different multiplicity of infection (MOI) (5, 10, 25, 50 and 100). The infection efficiency and fluorescence intensity were analyzed by inverted fluorescent microscopy. The clonal and proliferation ability, cell cycle and the mineralization potential were compared before and after transfection. Based on those mentioned above, we could evaluate the influence of infection on their biological characteristics. RESULTS AND CONCLUSION: Flow cytometry results showed that rat dental pulp stem cells expressed STRO-1 and CD146 but not CD34 or CD45. The dental pulp stem cells could differentiate into osteoblasts and adipocytes when cultured in specific medium for each lineage differentiation. The highest efficiency of infection and strongest fluorescence expression appeared at 48 hours of infection and MOI 50. There were no significant differences in growth ability, cell colony formation rate and cell cycle before and after transfection (P > 0.05). And the alkaline phosphatase expressed positively. Infection for 48 hours at MOI 50 is optimal for transfecting dental pulp stem cells with green fluorescent protein by a lentiviral vector, thereby providing reliable tracer method for the study of rat dental pulp stem cells 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Isolation and culture of mouse embryonic fibroblasts and preparation of feeder layers Hu San-qiang, Wang Yan-yan, Ma Yong-bin, Hu Jia-bo 2014, 18 (45):  7306-7311.  doi: 10.3969/j.issn.2095-4344.2014.45.017 Abstract ( 712 )   PDF (1182KB) ( 1215 )   Save OBJECTIVE: To establish the optimal method for isolation and culture of Kunming mouse embryonic fibroblasts, and to evaluate the feasibility of preparing feeder layers for culture of human embryonic stem cells. METHODS: Embryonic fibroblasts were isolated and cultured by different concentrations of trypsin from Kunming mouse fetuses in vitro. The biological characteristics and growth rule of mouse embryonic fibroblasts were investigated, and then the feeder layers for human embryonic stem cells culture were produced. The growth of human embryonic stem cells on the prepared feeder layer was tested. RESULTS AND CONCLUSION: The optimal fetal age for preparing Kunming mouse embryonic fibroblast feeder layer was 13.5 days. Kunming mouse embryonic fibroblasts at different concentrations grew well with high purity and active proliferation by trypsin digestion method. There was no significant difference in the survival rate of cells after cryopreservation for 2 weeks, 1 month, 3 months and 6 months. The cells were proliferative from the second to fourth passage and declined sharply after the fifth passage. Human embryonic stem cells which grew on Kunming mouse embryonic fibroblasts feeder layers were still to remain the typical undifferentiated morphology and were strongly positive for alkaline phosphatase and periodic acid-Schiff after long-term subculture. The mouse embryonic fibroblasts can be used as the stable and high-quality feeder cells for human embryonic stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Erythropoietin promotes endothelial progenitor cells proliferation depending on PI3k/Akt pathway Wu Hai-wei, Zhang Lei, Hu Ruo-yu, Li Hao, Jing Hua, Dong Guo-hua, Xu Biao, Li De-min 2014, 18 (45):  7312-7319.  doi: 10.3969/j.issn.2095-4344.2014.45.018 Abstract ( 307 )   PDF (715KB) ( 549 )   Save BACKGROUND: It has been proved that erythropoietin can promotes angiogenesis in injured tissue, which is closely related to the proliferation and differentiation of endothelial progenitor cells. However, the involved mechanism remains unclear yet. OBJECTIVE: To investigate the effect of erythropoietin on the function and activity of bone marrow-derived endothelial progenitor cells in mice, and to explore the signal pathway. METHODS: The endothelial progenitor cells from the bone marrow of mice were separated by means of density gradient centrifugation and then cultured. The cells were preconditioned by specific inhibitor of PI3K (LY294002), and were divided into the following groups: EGM-2 group, three erythropoietin preconditioned groups (the concentrations of erythropoietin in medium were 1, 5, 10 U/mL respectively), erythropoietin+LY group (10 U/mL erythropoietin and 10 mmol/L LY294002 in medium), LY group (10 mmol/L LY294002 in medium), dimethyl sulfoxide group (1 mL/L dimethyl sulfoxide in medium). The cell proliferation and apoptosis were evaluated by cell counting kit-8 and flow cytometry respectively. The contents of endothelial nitric oxide synthase and vascular endothelial growth factor in cell lysates were detected by the method of ELISA, and the expressions of Akt and p-Akt were by western blot assay. RESULTS AND CONCLUSION: Erythropoietin could promote the proliferation of endothelial progenitor cells in a dose-dependent manner, which was, however, completely inhibited by LY294002. The apoptosis rate in the erythropoietin preconditioned groups was significantly lower than that in the erythropoietin+LY group. The contents of endothelial nitric oxide synthase and vascular endothelial growth factor in cell lysates of LY group and erythropoietin+LY group were significantly lower than those in the erythropoietin groups. There was no difference in Akt expression found in each group, while the p-Akt expression in the erythropoietin+LY group was significantly lower than that in the erythropoietin groups. The above results reveal that erythropoietin can promote the proliferation of endothelial progenitor cells and decrease the cell apoptosis, which is depending on PI3K/Akt signal 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Adipogenic and tenogenic differentiation of tendon-derived stem cells isolated from an animal model of chronic tendinopathy in vitro Chen Hui, Lin Yu-cheng, Xu Hong-liang, Wang Chen, Rui Yun-feng 2014, 18 (45):  7320-7326.  doi: 10.3969/j.issn.2095-4344.2014.45.019 Abstract ( 393 )   PDF (739KB) ( 753 )   Save  BACKGROUND: Chronic tendinopathy is a tendon disorder extremely common in athletes and in the general population with repetitive strain injuries of tendons. The pathogenesis of tendinopathy remains unclear and hence treatment of tendinopathy is usually palliative. OBJECTIVE: To investigate the of adipogenic and tenogenic ability of patellar tendon-derived stem cells isolated from chronic tendinopathy and healthy rats in vitro. METHODS:Tendon-derived stem cells were isolated from patellar tendons of chronic tendinopathy and healthy rats respectively. The tendon-derived stem cells were cultured to the 3rd passage in complete culture medium, and cell morphology was observed. The cells were divided into adipogenic induction group and control group. Cells in the adipogenic induction group were cultured in adipogenic induction medium, while those in the control group cultured in complete culture medium. The ability of adipogenic differentiation between tendon-derived stem cells isolated from the tendon of chronic tendinopathy and healthy rats in vitro was examined by oil red O staining and quantification assay. The mRNA expressions of C/EBPα and PPARγ2 were detected by real-time quantitative PCR. When 70%-80% cells were confluent, the mRNA expressions of Col1a1, Scx, Tnmd and Dcn were also detected by real-time quantitative PCR. RESULTS AND CONCLUSION: At the third passage, slender spindle-shaped cells were seen in both two groups, but there was a little change in the cell morphology in the chronic tendinopathy group. Lipid droplets were formed after the cells were cultured in adipogenic induction medium for 21 days. This was not observed in the control group. We observed more oil red O-positive oil droplets in tendon-derived stem cells from the tendons of chronic tendinopathy rats than healthy rats. The difference between them was statistically significant (P=0.004). The results of real-time quantitative PCR showed that the mRNA expressions of C/EBPα and PPARγ2 in the tendon-derived stem cells from the tendons of chronic tendinopathy rats were significantly higher than those in tendon-derived stem cells from the tendons of healthy rats (P=0.004); the mRNA expressions of Col1a1, Scx, Tnmd and Dcn in the tendon-derived stem cells from the tendons of chronic tendinopathy rats were significantly lower than those in tendon-derived stem cells from the tendons of healthy rats (P=0.009). In conclusion, tendon-derived stem cells from chronic tendinopathy rats showed a higher ability of adipogenic differentiation, but a lower capacity of tenogenic differentiation compared to tendon-derived stem cells from healthy rats, which might contribute to better understand the pathogenesis of tendinopathy. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Chimerism of placenta-derived cells with maternal blood and umbilical cord blood cells Mo Zheng, Sheng Hong-xia, Han Zhong-chao, Xu Man, Tian Chong, Zhang Bin, Chen Hu 2014, 18 (45):  7327-7332.  doi: 10.3969/j.issn.2095-4344.2014.45.020 Abstract ( 384 )   PDF (493KB) ( 738 )   Save BACKGROUND: There are abundant cell populations in the placenta that attracts more and more attentions because of high content of CD34+ cells. It is expected to become a new source of hematopoietic stem cells for the treatment of hematologic diseases and other malignant diseases.   OBJECTIVE: To investigate the amount of cells derived from placenta, their colony forming ability, and their chimerism analysis. METHODS: Five placentas obtained from five healthy full-term cesarean women were treated with perfusion method and tissue digestion for the cell collection. Flow cytometry was used to detect the proportion of CD34+ cells in the placenta and cord blood, followed by the culture of cell colonies as well as regular observation of cell morphology and counting. PCR amplification with sequence-specific primers and sequence-specific oligonucleotide probes were used to examine HLA type of placenta, umbilical cord blood, and maternal peripheral blood; Short tandem repeat PCR was used for chimerism analysis. RESULTS AND CONCLUSION: There were more CD34+ cells in the placenta than in the umbilical cord blood. The placenta had good ability to form multiple colonies in vitro, and there were maternal source components in the placenta. It is concluded that the amount of cells in the placenta and their biological functions exhibit the potential use of placenta as a new source of hematopoietic stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Compliance of ischemic tissue to stem cells-derived vessels for secondary revascularization He Xiao-ming, Yang Yong, Wan Jia 2014, 18 (45):  7333-7336.  doi: 10.3969/j.issn.2095-4344.2014.45.021 Abstract ( 241 )   PDF (303KB) ( 532 )   Save BACKGROUND: To explore the methods for secondary limb revascularization after endovascular repair is one of the most urgent research topics. OBJECTIVE: To investigate the biological mechanism about the transplantation of autologous peripheral blood stem cells to lead and perfect the effect of ischemic tissue for secondary revascularization. METHODS: Forty-two patients with critical low limb ischemia were selected and treated with endovascular repair as first revascularization and transplantation of autologous peripheral blood stem cells as secondary revascularization. The secondary revascularization was carried out at 3 months after the first blood flow remodeling. The patients receiving secondary revascularization were followed for 4 years. RESULTS AND CONCLUSION: After transplantation of autologous peripheral blood stem cells, the patients complained that the rest pain of the lower leg was relieved obviously and intermittent claudication distance was significantly lengthened. Limb salvage rate was 100%. Skin temperature index was 1.6±0.3, transcutaneous oxygen pressure was (5.00±1.26) kPa, ankle-brachium index was 0.9±0.2, photoplethysmograpy index was 0.8±0.1, saturation of blood oxygen was (79.4±20.4)%, and digital subtraction angiography score was (1.3±0.2)points. After transplantation, local blood flow indicators of the low limbs were positively correlated to local symptom indicators of the limbs (0.6< r <0.7, P < 0.05). These results strongly suggest that the stem cells-derived neovascularization can offer effective and permanent blood flow perfusions to the ischemic tissue, and this biological effect play an important role in secondary revascularization of the ischemic tissue.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Transplantation of bone marrow mesenchymal stem cells in the treatment of bone nonunion following limb fractures: experimental results and conversion applications Yang Jun-li, Han Xia, Sun Ming-qi 2014, 18 (45):  7337-7341.  doi: 10.3969/j.issn.2095-4344.2014.45.022 Abstract ( 270 )   PDF (318KB) ( 413 )   Save BACKGROUND: As bone marrow mesenchymal stem cells can be differentiated into osteoblasts under certain induction conditions, autologous bone marrow mesenchymal stem cells can be implanted into the bone nonunion site of bone fracture. This new technology garners increasing attention of orthopedic clinicians. OBJECTIVE: To summarize the clinical efficacy of transplantation of bone marrow mesenchymal stem cells in the treatment of bone nonunion of limb fractures. METHODS: A computer-based search of Foreign Medical Journal Full-Text Service and CNKI databases was performed for articles related to bone marrow mesenchymal stem cells for treatment of bone nonunion of limb fractures published from 1998 to 2014 using the keywords of “bone marrow stem cells (BMSCs), stem cell transplantation (SCT), nonunions, tissue engineering” in English and Chinese, respectively. Literatures with repetitive content and lack of originality were excluded. A total of 36 literatures were obtained for further analysis. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells are transplanted into the end of bone nonunion, and then can be induced to differentiate into osteoblasts to repair bone nonunion and bone defects,laying a theoretical basis for clinical application. Bone marrow mesenchymal stem cells can repair bone defects, which provides an effective method and material to promote fracture healing. Transplantation of bone marrow mesenchymal stem cells is safe and effective for treatment of bone nonunion of limb fracture.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Application of mesenchymal stem cells in articular cartilage tissue engineering Zhang Yong-xing, Zhao Qing-hua 2014, 18 (45):  7342-7347.  doi: 10.3969/j.issn.2095-4344.2014.45.023 Abstract ( 468 )   PDF (364KB) ( 527 )   Save  BACKGROUND: The development of tissue engineering techniques provides new methods and ideas for the repair and functional reconstruction of articular cartilage defects. OBJECTIVE: To summarize the research progress in the application of mesenchymal stem cells, as seed cells, in articular cartilage tissue engineering. METHODS: A computer-based retrieval was performed to search articles describing articular cartilage tissue engineering and mesenchymal stem cells published between January 1st, 2000 and September 30th, 2014 in PubMed database with the key words of “articular cartilage defects; cartilage tissue engineering; mesenchymal stem cells” in English. Seventy articles were retrieved initially, and only 49 were included in further analysis. RESULTS AND CONCLUSION: The ability of the articular cartilage for defect self-repair is limited, and current  clinical treatments cannot be satisfactory. Development of tissue engineering provides a new idea for problem-solving. In the selection of seed cells, chondrocyte is limited to dedifferentiate, and embryonic stem cell is restricted by ethical, legal and other aspects. Autologous mesenchymal stem cells, which are easy to be amplified and exhibit excellent cartilage differentiation potential, have gained widespread attention. But there is still some controversy on the current application of tissue engineering techniques for repair of articular cartilage defects, including a certain gap between the long-term effects and the clinical applications. So the effect of mesenchymal stem cells on biological structure and mechanical function still needs 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Stem cell transplantation improves bone mass: research progress and application prospects  Cao De-yu, Li Feng, Qi Bao-chang, Gu Qun, Wang Chun-li, Tao Shu-qing 2014, 18 (45):  7348-7352.  doi: 10.3969/j.issn.2095-4344.2014.45.024 Abstract ( 283 )   PDF (596KB) ( 490 )   Save BACKGROUND: Studies have shown that bone loss can lead to a series of diseases, such as osteoporotic fractures, thus seeking to increase bone mass has become a goal of the majority of researchers. OBJECTIVE: To summarize the current studies of improving bone mass by using stem cell transplantation, hoping to the extensive application of stem cell transplantation in the clinical treatment of osteoporosis as early as possible. METHODS: A computer-based search of PubMed and CNKI was performed by the first author to retrieve articles relevant to stem cell therapy for osteoporosis published from January 1997 to October 2014. The keywords were “to improve bone mass, regenerative medicine, bone marrow mesenchymal stem cell transplantation, stem cell therapy” in Chinese and English, respectively, which appeared in the title, abstract or keywords. Articles published recently or in authoritative journals were preferred, and finally 28 articles were included in result analysis. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells which are isolated and cultured easily can proliferate rapidly and have multi-lineage differentiation potential. Studies have shown that the osteogenic differentiation of bone marrow mesenchymal stem cells can really improve bone mass, and obtain more achievements in the treatment of orthopedic disorders. This new cell therapy can help to accelerate bone healing and reduce treatment time, offering a new therapeutic choice for orthopedic surgery, plastic surgery, oral and maxillofacial surgery, and therefore, it has broad application prospects. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Endothelial progenitor cells in fracture healing: problems about culture and transplantation Meng Xin-min, Sun Xiao-lei, Ma Jian-xiong, Ma Xin-long 2014, 18 (45):  7353-7357.  doi: 10.3969/j.issn.2095-4344.2014.45.025 Abstract ( 258 )   PDF (293KB) ( 382 )   Save BACKGROUND: With the development of biochemistry and cell biology, fracture has being study deeper, blood supply has been known to be an important factor influencing the fracture healing. Endothelial progenitor cells with good ability of angiogenesis will have a good clinical prospect in fracture healing.   OBJECTIVE: To review the recent research of endothelial progenitor cells in fracture healing. METHODS: A computer-based online search of CNKI, Wanfang, PubMed databases was performed to collect articles published between 1980 and 2014 with the key words “endothelial progenitor cell, fracture, neovascularization, angiogenesis” in Chinese and English. A total of 48 articles addressing endothelial progenitor cell for angiogenesis in fracture healing were included in result analysis. RESULTS AND CONCLUSION: Increasing evidence has shown that endothelial progenitor cells have great ability of neovascularzition and angiogenesis. Endothelial progenitor cells used in tissue engineering scaffolds can promote the survival rate of scaffolds in vivo, which is appropriate to a great part of delayed union and nonunion patients. However, the large-scale treatment with endothelial progenitor cells still has many problems, such as isolation, culture and amplification of endothelial progenitor cells in vitro, the number of transplanted cells and selection of scaffolds for transplanted cells, which need further research. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Research progress of induced pluripotent stem cells and its bottleneck Ma Xun, Qin Jie, Xu Yu-ming 2014, 18 (45):  7358-7363.  doi: 10.3969/j.issn.2095-4344.2014.45.026 Abstract ( 739 )   PDF (349KB) ( 934 )   Save BACKGROUND: With the development of the research, induced pluripotent stem cells are applied to the build of disease model, drug screening, regenerative medicine, and many other research fields, and have made significant achievements, especially in the study of nervous system diseases.   OBJECTIVE: To summarize the recent development of induced pluripotent stem cells and to raise problems and prospects based on the latest research in this field. METHODS: The first author searched the PubMed database for articles about the induced pluripotent stem cells, including reviews, clinical research and basic research, published from January 2006 to September 2014. The keywords were “iPS, induced pluripotent stem cell”, and finally 60 articles were included in result analysis. RESULTS AND CONCLUSION: Induced pluripotent stem cell research continues to make breakthrough from its discovery by Yamanaka’s team in 2006 to winning Nobel Prize in 2012. Induced pluripotent stem cell research has broad prospects in the disease model construction, drug screening and regenerative medicine. Currently, problems such as reprogramming methods, cell stability, and clinical transformation still need to be solved, and further researches are necessary. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Stem cell transplantation for human diseases: current status and progress in basic research Tu Xue-song 2014, 18 (45):  7364-7369.  doi: 10.3969/j.issn.2095-4344.2014.45.027 Abstract ( 271 )   PDF (358KB) ( 471 )   Save BACKGROUND: With the development of stem cell culture technology, stem cell transplantation has been a research hotspot in the biological cytology. OBJECTIVE: To review the current situation and progress of basic research on stem cell transplantation for human refractory diseases. METHODS: A computer-based search of CNKI, VIP, Wanfang and PubMed databases was performed for articles related to animal experiments of stem cell transplantation published from January 2008 to October 2014. The key words were “induced pluripotent stem cells, neural stem cells, top-up between stem cells, transplantation” in Chinese and English in the title and abstract. Papers published in authoritative journals or recently were preferred. Totally 113 articles were searched initially, and only 50 articles were included in result analysis. RESULTS AND CONCLUSION: With the progress of stem cell culture technology, basic research on the stem cell transplantation for human diseases have been promoted and carried out, and these researches have achieved phased outcomes that provide conditions for more extensive and in-depth fundamental research. It is believed that with the development of basic research on stem cell transplantation to solve a series of difficult problems, stem cell transplantation will bring hope for treatment of human disease, especially for treatment of refractory disease. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Coculture of human umbilical vein endothelial cells and bone marrow mesenchymal stem cells Luo Shu-ping, Du Yu-ting, Bai Ju 2014, 18 (45):  7370-7374.  doi: 10.3969/j.issn.2095-4344.2014.45.028 Abstract ( 352 )   PDF (323KB) ( 542 )   Save BACKGROUND: Coculture of bone marrow mesenchymal stem cells and human umbilical vein endothelial cells can improve both osteogenic and angiogenic outcomes and provide a promising strategy for bone tissue engineering and osteanagenesis. OBJECTIVE: To summarize recent researches and related progresses in coculture of human umbilical vein endothelial cells and bone marrow mesenchymal stem cells. METHODS: A computer-based online search of CNKI database from January 2000 to March 2012, PubMed database and Web of Knowledge database from January 1980 to March 2012, was performed with the keywords of “human umbilical vein endothelial cells, bone mesenchymal stem cells, coculture, tissue engineering” both in Chinese and English. A total of 135 articles were screened out, 103 of them were excluded due to unrelated study objective and repeated contents, and finally 32 articles were involved in further analysis. RESULTS AND CONCLUSION: At present, studies on coculture of bone marrow mesenchymal stem cells and human umbilical vein endothelial cells mainly focus on mimicking in vivo environments, the interactions between cells, and the influence of different cell ratios and culture media. Most of these researches play important roles in bone tissue engineering and bone regeneration therapy, but the mechanism of action and concrete regulation in vivo between bone marrow mesenchymal stem cells and human umbilical vein endothelial cells still need further research and analysis. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Isolation methods of mesenchymal stem cells Cai Jin-hong, Lin Chun-bo, Yang Yuan 2014, 18 (45):  7375-7380.  doi: 10.3969/j.issn.2095-4344.2014.45.029 Abstract ( 257 )   PDF (382KB) ( 1193 )   Save BACKGROUND: Mesenchymal stem cells as potential seeded cells have been widely used in tissue engineering and clinic therapy; thus, the precise, safe, effective isolation of mesenchymal stem cells is the particular important premise to build culture system. OBJECTIVE: To review the methods of isolating mesenchymal stem cells and to compare the merit and demerit of different methods, thereby providing theoretical basis for safe and high-effective isolation of mesenchymal stem cells. METHODS: A computer-based online research of CNKI and PubMed databases was performed to collect articles, which included reviews, clinical trials and experiments, published between 1965 and 2014 with the key words of “mesenchymal stem cells (MSCs), isolation methods” in Chinese and English. A total of 52 articles were included according inclusion and exclusion criteria RESULTS AND CONCLUSION: (1) The whole bone marrow culture method can derive a mass of mesenchymal stem cells, which need to be purified. (2) The density gradient centrifugation method which uses the media with the density of 1.073 g/mL can be used to harvest more purified cells. (3) The tissue digestion method is suitable for digestion and isolation of adipose tissue and umbilical cord tissue. Type II collagenase digestion is better, but they are both limited by a high demand for operative techniques. (4) Immunomagnetic bead separation is appropriate to study the biological characteristics of a kind of subpopulation of mesenchymal stem cells which express special surface markers. (5) The combination method is also an optimal way. (6) Some new methods limited by few dates require 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 全文链接： Figures and Tables | References | Related Articles | Metrics