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28 November 2018, Volume 22 Issue 33 Previous Issue    Next Issue

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Effects of adipose-derived mesenchymal stem cells on fibroblast proliferation in mouse scleroderma lesions and underlying mechanisms Wang Wei, Huang Yu-cheng, Chen Yan-hui 2018, 22 (33):  5249-5254.  doi: 10.3969/j.issn.2095-4344.0657 Abstract ( 335 )   PDF (762KB) ( 183 )   Save BACKGROUND: Adipose-derived mesenchymal stem cells (ADSCs) maintain the potential of multidirectional differentiation during long-term culture in vitro, and can secrete various anti-fibrotic cytokines or growth factors that are involved in regulating fibroblast proliferation and inhibiting collagen deposition, and thereby exert anti-fibrotic effects. OBJECTIVE: To investigate the effects of ADSCs on the proliferation and apoptosis of fibroblasts in scleroderma mice and to explore the underlying mechanisms. METHODS: Fibroblasts were isolated and purified from the skin tissue of scleroderma mice. After passage, the cells were divided into five groups according to different treatment methods: control group, ADSCs intervention group, Wnt inhibitor HY-15597 treatment group (Wnt inhibitor group), Wnt activator SB-216763 treatment group (Wnt activator group), ADSCs+SB-216763 treatment group (combined group). Cell proliferation rate and apoptosis rate were detected using MTT and flow cytometry. The levels of collagen type I, collagen type III, and Wnt signaling pathway related proteins were detected and compared. RESULTS AND CONCLUSION: Compared with the control group, the proliferation rate of fibroblasts in the ADSCs intervention group and Wnt inhibitor group decreased and the apoptotic rate of fibroblasts increased significantly, while the proliferation rate increased and the apoptosis rate decreased in the Wnt activator group in a time-dependent manner (P < 0.05). Compared with the Wnt activator group, the proliferation rate of fibroblasts in the combined group was significantly lowered and the apoptotic rate was significantly increased (P < 0.05). Compared with the control group, the expression of hydroxyproline, collagen type I, collagen type III, Wnt2, Wnt3a, and β-catenin in the fibroblasts of ADSCs intervention group and Wnt inhibitor group were significantly decreased, while those in the Wnt activator group were significantly increased (P < 0.05). The levels of hydroxyproline, collagen type I, collagen type III, Wnt2, Wnt3a, and β-catenin in the combined group were significantly lower than those in the Wnt activator group (P < 0.05). The overall results indicate that ADSCs can inhibit the proliferation and collagen deposition of fibroblasts in the lesion tissue of mouse scleroderma models by inhibition of the Wnt/β-catenin 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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Feasibility of mixed human AB serum to expand human umbilical cord mesenchymal stem cells in vitro Hou Ping, Li Xiao-feng, Li Wei-wei, Li Jian-ping 2018, 22 (33):  5255-5258.  doi: 10.3969/j.issn.2095-4344.0658 Abstract ( 388 )   PDF (621KB) ( 259 )   Save BACKGROUND: At present, human umbilical cord mesenchymal stem cells (HUMSCs) are mainly cultured in a medium containing 10% fetal bovine serum or a combination of various cytokines or cultured in commercial serum-free medium, but there is no low-cost, relatively safe standardized culture system applicable to the clinical utility. OBJECTIVE: To explore the effective expansion of HUMSCs by mixed human AB serum. METHODS: HUMSCs were isolated by tissue explant method, and then cultured in the basal medium containing 10% mixed human AB serum or in the serum-free medium. Differences between the two culture systems were then detected, involving the morphological characteristics, proliferative capacity and cell cycle. RESULTS AND CONCLUSION: There was no significant difference in cell morphology between the two culture systems, and all the cultured cells were long shuttle-shaped. However, HUMSCs cultured with human AB serum had stronger adhesion. Flow cytometry results showed that HUMSCs highly expressed CD73 and CD90, but lowly expressed CD45 and HLA-DR, and the expression levels had no significant difference in both culture systems (P > 0.05). The cell cycle results showed that most cells were in G0-G1 and S phases, and the DNA index and coefficient of variation values in both culture systems were within the normal range. To conclude, the mixed human AB serum is considered to be a human-derived serum of clinical grade for safe use in the HUMSCs culture medium.  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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Viability of bone marrow mesenchymal stem cells in inflammation-induced ischemia-reperfusion environment Zhang Ni, Chen Lan-ying, Li Xue-liang, Guan Zi-yi, Fang Cong, Zhou Meng-jing, Luo Ying-ying, Liu Rong-hua 2018, 22 (33):  5259-5267.  doi: 10.3969/j.issn.2095-4344.0659 Abstract ( 428 )   PDF (1341KB) ( 149 )   Save BACKGROUND: Local tissue ischemia-hypoxia and inflammatory microenvironment mainly contribute to the low survival rate of bone marrow mesenchymal stem cells. OBJECTIVE: To study the effect of ischemia-hypoxia on viability of bone marrow mesenchymal stem cells in inflammatory environment and to explore the relationship between cell viability and inflammatory factors. METHODS: The whole bone marrow culture method was used to separate and culture bone marrow mesenchymal stem cells and flow cytometry was used for cell identification. The third passage cells were treated by oxygen-glucose deprivation (OGD) preconditioning for different periods (6, 12 and 24 hours) as well as inflammatory induction (OGD, OGD+20 μg/L tumor necrosis factor alpha (TNF-α), OGD+100 μg/L lipopolysaccharide (LPS)). The treated cells were observed under inverted microscope. Cell viability was tested by MTT assay and flow cytometry. The levels of interleukin-6, interleukin-10, and TNF-α in the cell supernatant were measured by ELISA and the mRNA expression levels of interleukin-6, interleukin-10, TNF-α and nuclear factor-κB in the cells were detected by RT-PCR. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells with a purity of 95% or more were successfully isolated and cultured. MTT and flow cytometry results showed that the cell viability was significantly increased after 6 and 12 hours of OGD, but was significantly decreased after 24 hours of OGD. The viability of bone marrow mesenchymal stem cells decreased after 6, 12, and 24 hours of OGD under the induction of TNF-α and lipopolysaccharide. The results of ELISA and RT-PCR showed that the levels of interleukin-6 and interleukin-10 in the cell supernatant were lowered after 6 and 12 hours of OGD and the gene expression of interleukin-6, interleukin-10, TNF-α, and nuclear factor-κB was lower compared with normal conditions. The levels of interleukin-6 and TNF-α in the cell supernatant increased significantly and the expression of interleukin-6, interleukin-10, TNF-α and nuclear factor-κB genes increased significantly after 6, 12 hours of OGD under TNF-α and LPS induction compared with simple OGD. Therefore, under the induction of TNF-α and lipopolysaccharide, the viability of bone marrow mesenchymal stem cells is affected by the expression and secretion of inflammatory factors. With the increase of the expression and secretion of inflammatory factors, the viability of bone marrow mesenchymal stem cells shows a decreasing trend.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of recombinant human granulocyte colony-stimulating factor on CXC chemokine receptor 4 expression and chemotaxis in bone marrow microenvironment Wang Jin, Niu Ben, Ma Xiao-rong, Zhang Wang-gang 2018, 22 (33):  5268-5273.  doi: 10.3969/j.issn.2095-4344.0648 Abstract ( 338 )   PDF (708KB) ( 156 )   Save BACKGROUND: The bone marrow microenvironment can mediate drug resistance and immune escape of leukemia cells through cell adhesion molecules. These adhesion molecules as new targets can be expected to break the drug resistance mechanism of leukemia cells by reducing their expression levels and functions. OBJECTIVE: To observe the effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on CXC chemokine receptor 4 (CXCR4) expression and chemotaxis in acute leukemia cells, as well as the level of stromal cell-derived factor-1 (SDF-1) in the bone marrow and peripheral blood CD4+CD25+Foxp3+ Treg cells. METHODS: (1) WEHI-3 cells were co-cultured with rhG-CSF for 6, 12, 18 and 24 hours, and the expression of CXCR4 was detected by flow cytometry. The cell migration was checked by Transwell assay. Healthy Balb/C mice were subcutaneously injected with rhG-CSF. The proportion of Treg cells in the peripheral blood and spleen was detected also by flow cytometry. ELISA was used to detect the level of SDF-1 in the bone marrow.  RESULTS AND CONCLUSION: rhG-CSF significantly reduced the CXCR4 expression on the WEHI-3 cell surface, as well as the chemotactic ability of SDF-1 (P < 0.05). The level of SDF-1 in the bone marrow of healthy mice treated with rhG-CSF was significantly lower than that of the control group (P < 0.05), and the proportion of Treg cells in the peripheral blood and spleen was significantly increased (P < 0.05). In conclusion, rhG-CSF can down-regulate CXCR4 expression on the surface of leukemia cells, reduce SDF-1 level in the bone marrow and mobilize the Treg cells into the peripheral blood. These changes might ultimately break the immune escape and drug resistance under the bone marrow microenvironment, thereby improving the effect of immunotherapy against leukemia.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Transplantation of adipose-derived stem cells combined with collagen bioengineering scaffold upregulates vascular endothelial growth factor expression in rats with chronic refractory wound Zhuang Jing, Yang Yu, Ding Li, Zheng Qing-jian 2018, 22 (33):  5274-5280.  doi: 10.3969/j.issn.2095-4344.0660 Abstract ( 300 )   PDF (977KB) ( 395 )   Save BACKGROUND: Bioengineering scaffolds are commonly used in the treatment of chronic refractory wounds and have achieved some effects, but there are also some problems. Adipose-derived stem cells (ADSCs) as a new method for the treatment of chronic refractory wounds have its unique efficacy and advantages. However, the efficacy and mechanism of ADSCs combined with bioengineering scaffolds in the treatment of chronic refractory wounds are not yet clear. OBJECTIVE: To study the effect of ADSCs combined with autologous collagen biomaterial scaffold transplantation on the expression of vascular endothelial growth factor (VEGF) during the healing of chronic refractory wounds. METHODS: Thirty-one Sprague-Dawley rats were purchased from Shanghai Slack Laboratory Animals Co., Ltd. Adipose tissues from the groin of a rat were harvested, and the primary ADSCs were isolated and cultured in vitro. The remaining 30 rats were randomly divided into normal control group, model group, collagen biomaterial scaffold group (Plenac group), autologous ADSCs transplantation group (ADSCs group) and autologous ADSCs + collagen biomaterial scaffold group (ADSCs+Plenac group), 6 rats in each group. In the normal control group, full-thickness skin wounds in the central and upper part of the bilateral lumbar vertebrae were made, followed by daily routine dressing. In the other groups two chronic refractory wounds were made in the central and upper part of the bilateral lumbar vertebrae followed by local injection of hydrocortisone acetate. Model group were given daily routine treatment; Plenac group treated with fibrin scaffold to cover the wound, followed by daily routine dressing; ADSCs group treated with autologous ADSCs transplantation followed by daily routine dressing; ADSCs+Plenac group received autologous ADSCs transplantation and bioengineering scaffold implantation to cover the wound, followed by daily routine dressing. After 7 days of intervention, all the rats were executed. The following procedures were subsequently carried out: observing wound situation, measuring wound size, detecting the expression of VEGF by immunohistochemical method, detecting the expression of VEGF protein using western blot assay, and detecting the expression of VEGF mRNA by qPCR.  RESULTS AND CONCLUSION: (1) The wound area in the control group was smaller than that in the other groups (P < 0.05), while the wound area in the ADSCs+Plenac group was also smaller than that in the model, Plenac and ADSCs groups (P < 0.05). (2) The expression of VEGF at protein and mRNA levels was higher in the control group compared with the other groups (P < 0.05), while the protein and mRNA expression of VEGF in the ADSCs+Plenac group was significantly higher than that in the model, Plenac and ADSCs group (P < 0.05). To conclude, ADSCs combined with the collagen bioengineering scaffold can upregulate VEGF expression in wound healing, and promote healing of wound healing. 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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Morphological characteristics of bone marrow mesenchymal stem cells differentiating into chondrocytes in vitro Wang Zhi-cong, Liu Yue-hong, Zhou Qing, Chen Xi, Zhou Yu, Sun Hui-jun, Liu Mo-zhen 2018, 22 (33):  5281-5285.  doi: 10.3969/j.issn.2095-4344.0661 Abstract ( 360 )   PDF (786KB) ( 163 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) can compact to form aggregates during chondrogenic differentiation, but whether the formation of aggregation is a prerequisite for chondrogenic induction is not yet documented. OBJECTIVE: To observe the morphological characteristics of BMSCs differentiating into chondrocytes. METHODS: BMSCs were cultured in vitro and passage 6 cells were used for the following experiments. Cells were induced in serum-free LG-DMEM medium as control group or in complete chondrogenic induction medium containing transforming growth factor β3 as induction group for 14 days. Cellular morphology during differentiation process was observed by inverted microscope. Chondrocyte-specific markers, including type II collagen, aggrecan and chondrogenic transcription factor SOX9, were evaluated by immunofluorescence, toluidine blue staining and quantitative RT-PCR. RESULTS AND CONCLUSION: Compared with the control group, the cells began to lose the typical morphology at day 3 of chondrogenic induction, and then compacted to form mono-layered aggregates and multi-layered aggregates at days 7 and 14, respectively. The aggregates were positive for immunofluorescence and toluidine blue staining, while no staining was detected in the single cells. qRT-PCR results also confirmed the mRNA expression of type II collagen, aggrecan and SOX9 in the cells after chondrogenic induction. Therefore, these abovementioned results suggest that the formation of aggregation in BMSCs can be one of morphological markers of chondrogenic differentiation.  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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Hepatic differentiation of bone marrow mesenchymal stem cells induced by additional growth factors inhibits chronic liver fibrosis Xiang Jun-xi, Liu Peng, Yang Li-fei, Su Jing-bo, Zhang Xu-feng, Liu Xue-min, Lü Yi 2018, 22 (33):  5286-5291.  doi: 10.3969/j.issn.2095-4344.0662 Abstract ( 322 )   PDF (898KB) ( 195 )   Save BACKGROUND: The use of bone marrow mesenchymal stem cells to improve hepatic fibrosis or cirrhosis is still controversial. Whether the effect of bone marrow mesenchymal stem cell transplantation for chronic liver fibrosis is related to the degree of differentiation remains to be further studied. OBJECTIVE: To explore the effect of hepatic-differentiated bone marrow mesenchymal stem cell transplantation in the treatment of chronic liver fibrosis. METHODS: Rat bone marrow mesenchymal stem cells were separated, cultured and purified by differential adherent culture. A 3-step protocol including sequential addition of growth factors, cytokines and hormones was used to induce the bone marrow mesenchymal stem cells to differentiate into hepatocyte-like cells. An animal model of chronic liver fibrosis was constructed by using tetrachloride. The bone marrow mesenchymal stem cells or hepatic-like cells were then implanted through the tail vein injection. The survival rate, liver fibrosis degree and liver function of rats were compared between groups. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells had the potential of differentiation in vitro and successfully differentiated into hepatic-like cells. After transplantation of hepatocyte-like cells, the 4-week survival rate of rats increased to 66.7%, the degree of liver fibrosis was significantly reduced to (14.69±0.37)%, the albumin level increased to (29.4±0.7) mg/L, and the level of glutamate transaminase decreased to (347.0±11.3) IU/L, which were significantly improved as compared with the model group and bone marrow mesenchymal stem cell transplantation group. To conclude, transplantation of hepatic-differentiated bone marrow mesenchymal stem cells can improve the liver function, reduce liver fibrosis and improve the survival rate of chronic liver fibrosis rats.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Biological characteristics of human placental mesenchymal stem cells isolated using two culture methods: a comparative study Zhang Fei, Wang Wu, Li Hui-juan, Liu Ya-fei, Zhang Bao-gang, Pei Fu-qiang, Shao Qing-wei, Wu Zhong-yan 2018, 22 (33):  5292-5296.  doi: 10.3969/j.issn.2095-4344.0644 Abstract ( 295 )   PDF (714KB) ( 180 )   Save BACKGROUND: Placental mesenchymal stem cells can provide ideal seed cells for cell therapy, but trypsin cold digestion has not been used for cell culture. Therefore, we explored this method and compared it with the tissue explant method to screen a more convenient and more successful culture method. OBJECTIVE: To observe the biological features of human placental mesenchymal stem cells isolated by trypsin cold digestion and tissue explant method. METHODS: We isolated, purified, and subcultured mesenchymal stem cells (n=6) from human placenta by using trypsin cold digestion method and tissue explant method. The first appearance time and primary culture period of placental mesenchymal stem cells isolated using two methods were recorded. The growth curves of passage 3 cells from two culture methods were generated. Flow cytometry was used to analyze the expression of surface markers of passage 3 cells isolated using the two culture methods, and to study the neural and adipogenic differentiation of the cells. RESULTS AND CONCLUSION: Placental mesenchymal stem cells could be obtained using both culture methods. Adherent cells in the trypsin cold digestion group appeared significantly earlier than those in the tissue explant group (P < 0.05), and the primary culture period was also significantly shorter in the trypsin cold digestion method (P < 0.05). The growth curve of passage 3 cells showed a higher cell number at plateau growth using trypsin cold digestion than tissue explant method (P < 0.05). The cells isolated using two culture methods were positive for CD29 and CD105, but negative for CD34 and CD45, indicating the same cell phenotypes. After the induction, placental mesenchymal stem cells cultured by both methods had the potential to differentiate into nerve cells and adipocytes. Therefore, both culture methods can produce placental mesenchymal stem cells, but trypsin cold digestion method is more suitable for the culture of this kind of cells as compared with the tissue explant 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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Co-transplantation of umbilical cord blood mesenchymal stem cells and haploidentical hematopoietic stem cells for treatment of severe aplastic anemia Ren Juan, Zhao Juan, Wang Xiao-ning 2018, 22 (33):  5297-5302.  doi: 10.3969/j.issn.2095-4344.0663 Abstract ( 361 )   PDF (624KB) ( 144 )   Save BACKGROUND: Nowadays the main therapy for severe aplastic anemia is immunosuppressive therapy or hematopoietic stem cell transplantation. With the development of hematopoietic stem cell transplantation technique, HLA-haploidentical hematopoietic stem cell transplantation is popularly used in the treatment of aplastic anemia. Mesenchymal stem cells as pluripotent stem cells can provide hematopoietic stem cells with ground substance and growth factors by interaction with hematopoietic stem cells in bone marrow. In addition, mesenchymal stem cells can inhibit the immunologic rejection and promote the immune reconstruction by regulating the immune function of the body. OBJECTIVE: To explore the efficacy and safety of co-transplantation of umbilical cord blood mesenchymal stem cells and HLA-haploidentical hematopoietic stem cells for the treatment of severe aplastic anemia. METHODS: From April 2015 to November 2017, six patients with severe aplastic anemia received fludarabine, cyclophosphamide and antithymocyte globilin as preconditioning before transplantation of HLA-haploidentical hematopoietic stem cells from the bone marrow and peripheral blood. Umbilical cord blood mesenchymal stem cells were infused before re-infusion of mobilized peripheral blood and bone marrow stem cells. Prophylaxis for graft-versus-host disease consisted of cyclosporine-A, short-course methotrexate as well as mycophenolate mofetil in patients. RESULTS AND CONCLUSION: One of six cases died of severe infection at +102 days after transplantation, with neutrophil engraftment at +48 days, and platelets were not engrafted until died. The remaining five cases were in disease-free survival with the median neutrophil engraftment at +11 days, and the median platelet engraftment at +21 days after transplantation. Grade I to II acute graft-versus-host disease was observed in the two of six patients, and cytomegalovirus viremia in three patients as well as epstein-barr virus infection in one patient. Co-transplantation of umbilical cord blood mesenchymal stem cells and haploidentical hematopoietic stem cells can be a choice for refractory severe aplastic anemia without HLA-matched related or unrelated donor, which is worth exploring in the future. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Human urine-derived stem cells transplantation for treatment of myocardial infarction in rats Sun He-yuan 2018, 22 (33):  5303-5308.  doi: 10.3969/j.issn.2095-4344.0664 Abstract ( 271 )   PDF (815KB) ( 90 )   Save BACKGROUND: In recent years, many studies have shown that bone marrow mesenchymal stem cells, adipose-derived stem cells, and cardiac stem cells can participate in myocardial repair, but little is reported on the use of human urine-derived stem cells in the treatment of myocardial infarction. OBJECTIVE: To investigate the effect of human urine-derived stem cell transplantation on the heart function of myocardial infarction rats. METHODS: The 20 of 64 Sprague-Dawley rats were selected as normal control group, and the remaining 44 rats were subjected to ligation of the left anterior descending coronary artery to make myocardial infarction models. The 40 model rats were successfully made and randomized into myocardial infarction group and cell transplantation group with 20 rats in each group. The rats in the myocardial infarction group were given injection of normal saline by tail vein, while those in the cell transplantation were given injection of human urine-derived stem cells labeled by CM-Dil by tail vein. The treatment was done once a day and lasted for 3 consecutive days. At 6 weeks after cell transplantation, the heart function of rats was examined by echocardiography, and the myocardial tissues of rats in each group were taken for CD34 immunohistochemical staining, hematoxylin-eosin staining, TUNEL staining and RT-PCR detection. RESULTS AND CONCLUSION: Compared with the myocardial infarction group, significantly improved heart function of rats, increased density of neonate blood capillaries, and decreased apoptotic rate were observed in the cell transplantation group (all P < 0.05). The CM-Dil positive cells were not observed in the normal control group and myocardial infarction group, while a great number of positive cells appeared in the cell transplantation group. Compared with the myocardial infarction group, Bcl-2 gene expression level in the cell transplantation group was significantly higher (P < 0.05), while the Caspase-3 gene expression level was lowered (P < 0.05). In conclusion, transplantation of human urine-derived stem cells can significantly improve the heart function of rats with acute myocardial infarction, and reduce cell apoptosis in the rat myocardium.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Bone defect repair with bone morphogenetic protein 14 transfected adipose-derived stem cells seeded onto a silk fibroin/hydroxyapatite scaffold Ding Tao, Tang Feng-lin, Lu Miao, Gu San-jun, Wang Jian-bing, Liu Hao 2018, 22 (33):  5309-5314.  doi: 10.3969/j.issn.2095-4344.0665 Abstract ( 342 )   PDF (1461KB) ( 185 )   Save BACKGROUND: In clinic, large bone defects are often difficult to repair naturally. Traditional methods including autologous and allogeneic bone grafts gradually show their limitations. However, the development of tissue engineering technology is expected to solve this problem. Transgenic technology is used to induce osteogenic induction of stem cells, and then induced cells are seeded onto artificial scaffold materials to construct tissue-engineered bone in vitro, which brings new hope for the repair of large-area bone defects. OBJECTIVE: To explore the biocompatibility of bone morphogenetic protein 14 (BMP14) transfected adipose-derived stem cells with silk fibroin (SF)/hydroxyapatite (HA) scaffolds and to evaluate the effect on repairing bone defects with this tissue-engineered bone. METHODS: Rat adipose-derived stem cells were isolated and cultured in vitro using collagenase digestion method. The third passage cells were transfected by lentivirus with BMP14 gene. Transfection efficiency was counted under fluorescent microscope. Meanwhile, the SF/HA scaffolds was prepared. Transfected cells were then seeded onto the SF/HA scaffold to prepare tissue-engineered bone. Biocompatibility of the prepared tissue-engineered bone was tested by scanning electron microscope and MTT method. Afterwards, the prepared tissue-engineered bone was implanted into the defect of the rat caudal vertebra as experimental group, and empty transfected cells with SF/HA scaffolds acted as control group. Remediation effect on bone defects was evaluated by X-ray and micro-CT examination. RESULTS AND CONCLUSION: BMP14 transfected adipose-derived stem cells adhered to the SF/HA scaffold and grew well. At 8 weeks after surgery, X-ray and micro-CT images showed that bone density of the intervertebral space obviously increased and a large number of calli developed in the experimental group, indicating a better recovery in bone continuity, while in the control group, the scaffold material incompletely absorbed was visible in the bone defect, and no obvious new bone tissues formed to fill in the defect. To conclude, these findings reveal that the SF/HA scaffold has good biocompatibility with BMP 14-transfected adipose-derived stem cells, and the composite artificial bone has good outcomes in the repair of bone defects.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Transplantation of human umbilical cord mesenchymal stem cells for mouse skin ulcer Li Mao, You Cheng-dong, Huang Wen 2018, 22 (33):  5315-5320.  doi: 10.3969/j.issn.2095-4344.0666 Abstract ( 271 )   PDF (997KB) ( 177 )   Save BACKGROUND: More assessments are required on the use of human umbilical cord mesenchymal stem cells in the treatment of skin ulcers. OBJECTIVE: To explore the feasibility of human umbilical cord mesenchymal stem cells to promote skin ulcer healing in mice. METHODS: Twenty C57 mice were randomized into control and treatment groups (n=10 per group). A longitudinal skin wound model was made on the back of the mouse with sterile scissors. The wound surface was about 2.5 cm×2.0 cm in size and the wound depth was about 5 mm. After modeling, 1 mL of human umbilical cord mesenchymal stem cells was injected into the wound in the treatment group, while the same volume of normal saline was given in the control group. The ulcer area of the two groups was measured every day, and the wound healing rate was calculated. The peri-ulcer tissue was taken at 0, 7, and 14 days after transplantation. The expression of vascular endothelial growth factor was detected by immunohistochemistry. The anti-nuclear antibody MAB1281 was detected by immunofluorescence staining to assess the residual rate of human umbilical cord mesenchymal stem cells around the ulcer. RESULTS AND CONCLUSION: The skin ulcer in the treatment group healed more quickly than that in the control group (P < 0.05). The expression of vascular endothelial growth factor was higher in the treatment group than the control group (P < 0.05). There were more residual human umbilical cord mesenchymal stem cells at the day of transplantation, while the number of residual cells gradually reduced with the healing of skin ulcer. These findings indicate that human umbilical cord mesenchymal stem cells maybe promote the neovascularization by paracrine approaches, and thus facilitate the healing of skin ulcer in 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 Figures and Tables | References | Related Articles | Metrics

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Calcitonin gene-related peptide regulates proliferation and osteogenic differentiation of synovial mesenchymal stem cells Deng Rui, Pan Fu-wen, Han Yao-guang 2018, 22 (33):  5321-5326.  doi: 10.3969/j.issn.2095-4344.0667 Abstract ( 345 )   PDF (724KB) ( 140 )   Save BACKGROUND: It has been proved that calcitonin gene-related peptide can promote the proliferation and osteogenic differentiation of synovial mesenchymal stem cells. However, little is reported on its specific mechanisms. OBJECTIVE: To investigate the effects of calcitonin gene-related peptide intervention on the proliferation and osteogenic differentiation of synovial mesenchymal stem cells, and to explore the role of Wnt pathway/β-catenin in this process.    METHODS: Synovial mesenchymal stem cells were isolated from synovial tissues of the rabbit knee joint by trypsin and collagenase I digestion. The third generation suspension of synovial mesenchymal stem cells was selected and cultured in four groups. The blank control group was routinely cultured without any intervention. In the experimental group, 10-8 mol/L calcitonin gene-related peptide was added. In the inhibitor group, 10-8 mol/L DKK-1, a Wnt signal pathway inhibitor, was added. In the combined intervention group, 10-8 mol/L calcitonin gene-related peptide and 10-8 mol/L DKK-1 were added. After 14 days of intervention, related indicator detections were performed. RESULTS AND CONCLUSION: (1) The expression of β-catenin protein in the inhibitor group was lower than that in the blank control group (P < 0.05); the expression of β-catenin protein in the experimental group was higher than that in the blank control group (P < 0.05); and the expression of β-catenin protein in the combined intervention group was higher than that in the inhibitor group (P < 0.05). (2) The proliferation of cells in the inhibitor group was slower than that in the other three groups at 3-9 days of culture (P < 0.05), while the proliferation of cells in the experimental group was always faster than that in the blank control group and combined intervention group at 5-9 days of culture (P < 0.05). (3) The activity of alkaline phosphatase, mineralized nodule area and mRNA expression of bone morphogenetic protein 2, Runx2, alkaline phosphatase and type I collagen in the experimental group were higher than those in the other three groups (P < 0.05). Compared with the blank control group, the inhibitor groups showed decreased alkaline phosphatase activity, reduced mineralized nodule area as well as reduced expression of bone morphogenetic protein 2, Runx2, alkaline phosphatase and type I collagen, while these indicators were higher in the combined intervention group than the inhibitor group (P < 0.05). To conclude, intervention with calcitonin gene-related peptide can promote the proliferation and osteogenic differentiation of synovial mesenchymal stem cells through the Wnt/β-catenin 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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Effects of intravenous transplantation of human umbilical cord blood mesenchymal stem cells on conversion of intra-and extra-cardiac monocyte-macrophages M1/M2 subtypes in mice with acute myocardial infarction Peng Yi, Chen Bing-quan, Zhao Ji-ling, Peng Zhi-yong, Xu Wei-fang, Yu Guo-long 2018, 22 (33):  5327-5332.  doi: 10.3969/j.issn.2095-4344.0643 Abstract ( 354 )   PDF (1085KB) ( 193 )   Save BACKGROUND: Studies have shown that transplantation of human umbilical cord blood mesenchyme stem cells (hUCB-MSCs) may reduce inflammation, improve cardiac function and participate in ischemic myocardial protection. However, there is still no final conclusion on the anti-inflammatory effect of hUC-MSCs. OBJECTIVE: To investigate the effects of intravenous transplantation of hUCB-MSCs on the conversion of intra- and extra-cardiac monocyte-macrophages subtypes LY-6C hi (M1)/LY-6 low (M2) and cardiac protection in mice with acute myocardial infarction (AMI). METHODS: Balb/C mice with AMI, aged 7-8 weeks, were randomly divided into experimental group (n=7) and control group (n=8). At 1 week after AMI, the experimental group was injected with 0.2 mL of saline containing 1×106 hUCB-MSCs via tail vein, and the control group was given 0.2 mL of saline with no cells. At 4 weeks after treatment, the M1/M2 ratio in the peripheral blood and spleen was measured by flow cytometry. Cardiac function was measured by echocardiography. Pathological changes of the myocardium, collagen sediment, apoptosis in myocardial cells, and myocardial macrophage M1/M2 cell count were detected by hematoxylin-eosin staining, Masson staining, TUNEL staining and immunofluorescence analysis, respectively. RESULTS AND CONCLUSION: Compared with the control group, monocyte-macrophages M1/M2 ratio in the peripheral blood, spleen and peri-infarcted myocardial tissue was lower in the experimental group (P < 0.05). Compared with the control group, cardiac functions were significantly improved in the experimental group presenting with lower left ventricular end-systolic diameter, left ventricular end-diastolic diameter as well as higher left ventricular ejection fraction (P < 0.05). Compared with the control group, the inflammatory cell count in the infarction area and surrounding tissue was significantly lower in the experimental group (P < 0.05). Compared with the control group, the amount of collagen sediment was significantly less in the experimental group (P < 0.05). Compared with the control group, the apoptosis index was lower in the experimental group, but there was no significant difference between the two groups (P > 0.05). These experimental findings verify that the intravenous transplantation of hUCB-MSCs can improve cardiac function of AMI mice by regulating systemic inflammatory responses and focal infarct lesions, which is one of the mechanisms by which hUCB-MSCs transplantation for myocardial infarction produces myocardial protection.  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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Expression of Toll-like receptor 4 in dental pulp stem cells under the stimulation of lipopolysaccharide and Streptococcus mutans Liu Ying, Gao Jie, Wu Bu-ling 2018, 22 (33):  5333-5337.  doi: 10.3969/j.issn.2095-4344.0668 Abstract ( 339 )   PDF (671KB) ( 200 )   Save BACKGROUND: During the process of deep caries, dental pulp stem cells (DPSCs) are induced to differentiate into odontoblast-like cells after bacterial stimulation, forming reparative dentin to protect pulp tissues. It is considered to be an important part of innate immune response of dental pulp. However, the role of Toll-like receptor 4 (TLR4) as the most important receptor of innate immune response is not clear. OBJECTIVE: To study the expression of TLR4 with the stimulation of lipopolysaccharide and Streptococcus mutans during deep caries. METHODS: Primary cultured DPSCs were stimulated by lipopolysaccharide and Streptococcus mutans extract for 24 hours. RT-PCR was then performed to test the expression of TLR4 mRNA. Western blot and immunocytochemical staining were performed to test the expression of TLR4 protein in DPSCs. RESULTS AND CONCLUSION: Stimulations with lipopolysaccharide and Streptococcus mutans extract significantly up-regulated the mRNA and protein expression of TLR4, indicating TLR4 is involved in the innate immune response of DPSCs in the case of deep caries. 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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Ginsenoside Rg1 protects against hydrogen peroxide induced damage to bone marrow-derived endothelial progenitor cells Jiang Wen-jie, Liang Xue-mei 2018, 22 (33):  5338-5343.  doi: 10.3969/j.issn.2095-4344.0646 Abstract ( 317 )   PDF (724KB) ( 128 )   Save BACKGROUND: Studies have confirmed that ginsenoside Rg1 has protective effects on vascular endothelial cells, but its underlying mechanism is still unclear. OBJECTIVE: To investigate the mechanism by which ginsenoside Rg1 protects against hydrogen peroxide induced injury of rat bone marrow-derived endothelial progenitor cells. METHODS: Endothelial progenitor cells derived from bone marrow of Sprague-Dawley rats were isolated and cultured by direct adherent method. The endothelial progenitor cells in logarithmic growth period were divided into three groups: control group, no intervention but conventional culture for 24 hours; cell injury group, cells were cultured in EMG-2 medium containing 100 μmol/L hydrogen peroxide for 24 hours; combined intervention group, cells were pretreated with ginsenoside Rg1 (64 μmol/L) for 2 hours, then cultured in EMG-2 medium containing 100 μmol/L hydrogen peroxide for 22 hours. Cell proliferation, apoptosis and migration were detected using cell counting kit-8, TUNEL, and cell scratch test, respectively. The expression of Akt, p-Akt, Bax, and Bcl-2 proteins was measured using western blot assay. The levels of intracellular superoxide dismutase, malondialdehyde and nitric oxide were determined using ELISA method. RESULTS AND CONCLUSION: (1) Compared with the cell injury group, the cell survival rate and cell migration ability were significantly increased, while the number of apoptotic cells reduced significantly in the combined intervention group (all P < 0.05). (2) Compared with the cell injury group, the levels of intracellular superoxide dismutase and nitric oxide were significantly increased, while the level of malondialdehyde was significantly reduced in the combined intervention group (all P < 0.05). (3) No significant difference in the expression of Akt protein was detected among the three groups, but the expression of p-Akt protein was significantly higher in the combined intervention group than the cell injury group (P < 0.05). Moreover, significantly increased Bcl-2 expression and decreased Bax expression were observed in the combined intervention group as compared with the cell injury group. In conclusion, ginsenoside Rg1 can antagonize hydrogen peroxide-induced damage to rat bone marrow-derived endothelial progenitor cells, increase cell viability, inhibit cell apoptosis, and reduce oxidative stress injury via the Erk 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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miR-124 facilitates the proliferation and differentiation of neural stem cells by inhibiting the Notch pathway He Pin, Guo Fu-qiang 2018, 22 (33):  5344-5349.  doi: 10.3969/j.issn.2095-4344.0669 Abstract ( 334 )   PDF (734KB) ( 300 )   Save BACKGROUND: How to quickly and efficiently obtain enough pure neurons in vitro is a huge challenge and problem faced by the scientists in the cell-transplantation field. OBJECTIVE: To explore whether miR-124 can influence the proliferation and differentiation of neural stem cells by regulating the Notch pathway. METHODS: The fetal rat neural stem cells were isolated and identified by immunofluorescence. RT-qPCR was used to detect the expression of miR-124 after 1, 2, 4, and 7 days of neural stem cell differentiation. After overexpression and interference with miR-124 via transient transfection, the expression of ki-67 protein was detected by MTT assay and immunoblotting. RT-qPCR and immunoblotting were used to detect β-tubulin III expression for determination of miR-124 effects on cell differentiation. RT-qPCR was used to detect the mRNA expression of Notch pathway-related proteins HES1, HEY2 and CCND1. RESULTS AND CONCLUSION: During the differentiation of neural stem cells into neurons, the expression of miR-124 was significantly upregulated. Moreover, miR-124 overexpression promoted the proliferation and differentiation of NSCs, and suppressed the expression of proteins in the Notch pathway. Silencing the miR-124 expression, however, achieved the opposite results. The above results suggest that miR-124 may promote the proliferation and differentiation of neural stem cells by inhibiting the Notch 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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Compatibility of nasal mucosa-derived ectomesenchymal stem cells and calcium phosphate cement in vitro Shi Wen-tao, Dai Yao, Chen Ping-bo, Bi Shi-qi, Lü De-ming, Lu Hao, Zhang Zhi-jian 2018, 22 (33):  5350-5355.  doi: 10.3969/j.issn.2095-4344.0670 Abstract ( 329 )   PDF (906KB) ( 207 )   Save BACKGROUND: The use of calcium phosphate cement (CPC) carrying mesenchymal stem cells for the treatment of bone defects has achieved satisfactory results. The in vitro compatibility of ectomesenchymal stem cells (EMSCs) derived from the nasal mucosa with CPC is unknown.  OBJECTIVE: To investigate the effect of calcium phosphate cement on the differentiation of rat nasal mucosa-derived EMSCs into osteoblasts and to further explore the biocompatibility of EMSCs and CPC.  METHODS: EMSCs were cultured, expanded in vitro and implanted onto the CPC surface or onto culture plates. Then, implanted EMSCs were induced to differentiate into osteoblasts for 14 days. Alkaline phosphatase activity assay was used to detect the alkaline phosphatase activity of EMSCs. The area of calcium nodules was determined by alizarin red staining. The expression level of osteogenesis-related proteins was detected by western blot assay and immunofluorescence method. Morphology of EMSCs on the CPC surface was dynamically observed using phalloidin staining. MTT assay was used to detect the proliferation of EMSCs on the CPC surface.  RESULTS AND CONCLUSION: After 14 days of osteogenic induction, there was higher alkaline phosphatase activity, larger calcium nodules and higher levels of osteogenesis-related proteins in the EMSCs implanted on the surface of CPC than those implanted on the culture plates. The CPC had a certain role in promoting the proliferation of EMSCs. To conclude, EMSCs and CPC scaffolds have good biocompatibility, and moreover, CPC can certainly promote the differentiation of EMSCs into osteoblasts.  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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Three kinds of mesenchymal stem cells: differentiating into neuron-like cells Yang Ying, Xu Qian, Wang Hui-ling, Yao Min-xiu 2018, 22 (33):  5356-5361.  doi: 10.3969/j.issn.2095-4344.0671 Abstract ( 328 )   PDF (617KB) ( 270 )   Save BACKGROUND: Stem cells, characterized with convenient collection, wide sources and great potential, have been developed in the cell therapy. The most commonly used stem cells include bone marrow mesenchymal stem cells, placental mesenchymal stem cells and umbilical cord blood mesenchymal stem cells. OBJECTIVE: To compare the biological changes in bone marrow mesenchymal stem cells, placental mesenchymal stem cells and umbilical cord blood mesenchymal stem cells after co-cultured with nerve cells. METHODS: Human bone marrow mesenchymal stem cells, human placental mesenchymal stem cells and human umbilical cord blood mesenchymal stem cells were isolated and cultured in vitro. The morphological changes of mesenchymal stem cells differentiated into nerve cells were observed by co-culture with Transwell culture plates. The expression of neuron-specific enolase was detected after induction. RESULTS AND CONCLUSION: In the co-culture system, these three kinds of mesenchymal stem cells gradually formed a star-like shape and were interconnected. Moreover, all the cells were positive for neuron-specific enolase. These findings reveal that the microenvironment provided by nerve cells can induce and promote the differentiation of three kinds of 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 Figures and Tables | References | Related Articles | Metrics

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The characteristics and significance of non-clonal chromosome aberrations in the in vitro expanded NK cells Li Yu-long, Huang Zhou-feng, Zhang Lei, Wu Cheng-ye, Cheng Wei, Dong Xiao-yan, Zhu Zun-min, Sun Kai 2018, 22 (33):  5362-5367.  doi: 10.3969/j.issn.2095-4344.0645 Abstract ( 335 )   PDF (703KB) ( 230 )   Save BACKGROUND: The significance of non-clonal chromosome aberrations (NCCAs) in in vitro expanded stem cells has been discussed in many studies; however, it is not well documented in the expanded NK cells. OBJECTIVE: To investigate the characteristics and significance of NCCAs in the in vitro expanded NK cells. METHODS: Peripheral blood mononuclear cells were isolated from patients with tumors and healthy controls. Artificial antigen presenting cells were used to stimulate the proliferation of NK cells. On the 7th day of culture, the cells were collected for phenotype identification and interferon-γ secretion test by flow cytometry and for cytogenetic analysis by the conventional G-banding technique. The difference in the frequency of NCCAs and interferon-γ secretion between the two groups and the correlation between NCCAs and interferon-γ secretion were studied. RESULTS AND CONCLUSION: NCCAs were found in 6 of 7 samples from the tumor patient group and 6 of 12 samples from the healthy controls. In the tumor patient group, 17 metaphases containing NCCAs were found out of 481 metaphases, and in the control group, 7 NCCA-containing metaphases were detected out of 785 metaphases [3.53% (17/481) vs. 0.89% (7/785), Z=-2.212, P=0.028]. Four patients with a history of chemoradiotherapy showed higher NCCA frequencies than those patients undergoing chemoradiotherapy (P=0.034) and the healthy controls (P=0.003). No difference was found between the newly diagnosed tumor patients and the healthy controls (P=0.819). The ratios of interferon-γ secreting NK cells in the two groups were not significantly different (P=0.371) and the correlation between the frequency of NCCAs and the interferon-γ secreting level was not identified. It was the first time that the characteristics of NCCAs in the artificial antigen presenting cells-expanded NK cells and the relationship between NCCAs and the NK cell function were preliminarily studied. The effect of NCCAs on the function and survival of NK cells in vitro and in vivo needs further investigations. 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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Optimal temperature and storage time span for cryopreservation of platelets Yang Zhen-yu, Li Zheng-fa, Fu Ling-mei, Zhu Xiang-ming 2018, 22 (33):  5368-5372.  doi: 10.3969/j.issn.2095-4344.0642 Abstract ( 623 )   PDF (595KB) ( 192 )   Save BACKGROUND: Clinical demand of fresh liquid platelets cannot be guaranteed because of the rapid increase of the need of platelets, especially for the specific blood types, emergency patients and remote areas. OBJECTIVE: To explore the optimal temperature and storage time span for deep cryopreservation of platelets in 5% dimethyl sulfoxide as well as the optimal usage time after melting of the frozen platelets. METHODS: Sixteen platelet samples filtered by the machine were randomly selected. Indexes as blood platelet count, mean platelet volume, platelet distribution width, CD42b, CD62p were separately tested before and 1, 3, 6, 12 months after cryopreservation at three different temperatures of -18, -40, and -80 ℃ as well as immediately, 1, 2, 4 hours after melting. RESULTS AND CONCLUSION: Significant changes were found in the expression of CD62p in the platelets at 6 and 12 months after cryopreservation at -80 ℃ and the expression of CD42b in the platelets at 6 months after cryopreservation at -80 ℃ (P < 0.05). The expression of CD42b and CD62p showed a significant difference in the platelets stored at -80 ℃ and at -18 and -40 ℃ for 6 months (P < 0.05). For the platelets stored at -80 ℃ for 6 months, the expression of CD42b and CD62p in the platelets at 4 hours after melting was significantly different from that immediately after melting (P < 0.05). These findings reveal that the lower temperature indicates the better cryopreservation of platelets using 5% dimethyl sulfoxide. The recommended storage conditions are -80 ℃ for 6 months; and the optimal usage time after melting is within 2 hours. 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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Mesenchymal stem cells for treating osteogenesis imperfecta: directly differentiating into functional cells or functioning via paracrine mechanism? Wang Zi-han, Liu Yi, Ju Ming-yan, Li Guang 2018, 22 (33):  5373-5378.  doi: 10.3969/j.issn.2095-4344.0672 Abstract ( 334 )   PDF (649KB) ( 245 )   Save BACKGROUND: Some limitations involving drug therapies and surgical treatment exist in the traditional treatment of osteogenesis imperfecta. In recent years, a growing number of scientists attempt to treat osteogenesis imperfecta by mesenchymal stem cells transplantation. OBJECTIVE: To summarize the mesenchymal stem cell treatment of osteogenesis imperfecta to guide relevant animal experiments, thereby promoting its clinical application. METHODS: A computer-based online search of PubMed between January 2000 and June 2018 was performed to search related articles with the keywords of “osteogenesis imperfecta, stem cells, mesenchymal stem cells, stem cells therapy, stem cells transplantation” in English. Literatures regarding mesenchymal stem cells for treatment of osteogenesis imperfecta were selected; in the same field, the articles published lately in authoritative journals were preferred. RESULTS AND CONCLUSION: Mesenchymal stem cells can directly differentiate into functional cells or exert paracrine effect to repair bone defects caused by osteogenesis imperfecta. The low immunogenicity and immunomodulatory capacity of mesenchymal stem cells ensure the transplantation safety in the treatment of osteogenesis imperfecta. Mesenchymal stem cells can fundamentally treat osteogenesis imperfecta, and do not produce adverse reactions that are unavoidable in the drug treatment, which have broad application prospects. Although there are currently no systematic criteria for the efficacy and safety of mesenchymal stem cells in the treatment of osteogenesis imperfecta, mesenchymal stem cell transplantation can circumvent the limitations of conventional drugs and surgical treatment. Further studies are expected to improve the treatment of osteogenesis imperfecta using mesenchymal stem cells and promote its clinical applications, as cell transplantation indeed benefits the patients.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Mesenchymal stem cells for cartilage repair in osteoarthritis Zhang Shuang, Liu Shi, Wang Yu-feng, Lü Chan, Li Bao-ling, Wang Ping, Cong Jun, Zhu Zhi-qiang 2018, 22 (33):  5379-5385.  doi: 10.3969/j.issn.2095-4344.0673 Abstract ( 279 )   PDF (684KB) ( 330 )   Save BACKGROUND: The degeneration of articular cartilage represents an ongoing challenge at the clinical and basic levels. Considering the restricted intrinsic capacity of resident chondrocytes to regenerate post injury, stem cell-based therapies have been proposed as a novel therapeutic approach for cartilage repair. Mesenchymal stem cells are pluripotent and immunosuppressive, which have become the first choice for tissue engineering research. OBJECTIVE: To summarize the application of mesenchymal stem cells derived from different tissues in cartilage repair in recent years. METHODS: With “mesenchymal stem cells, cartilage damage repair, osteoarthritis” as retrieval words, we searched relevant articles regarding mesenchymal stem cells for cartilage repair in PubMed, Web of Science and CNKI. Initially 168 articles were retrieved, and 59 eligible articles (57 in English and 2 in Chinese) were included in final analysis. RESULTS AND CONCLUSION: Mesenchymal stem cells with self-renewal ability can differentiate into a variety of mesodermal lineage cells, such as osteoblasts, chondrocytes, adipocytes, cardiomyocytes and vascular cells. The immunomodulatory and regenerative capacity of mesenchymal stem cells can regulate the imbalance of anabolism and catabolism in the osteoarthritic joint. Although a large number of mesenchymal stem cells have been used for cartilage repair in clinical practice, the mechanism underlying cartilage repair with mesenchymal stem cells is still unclear. Moreover, the therapeutic effect of mesenchymal stem cells is determined by cell sources, the number of transplanted cells and the way of injection. All of these problems need further explorations.  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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Mechanism of stretch-activated ion channels in mechanotransduction of mesenchymal stem cells Su Hao, Jia Xiao-ling 2018, 22 (33):  5386-5392.  doi: 10.3969/j.issn.2095-4344.0674 Abstract ( 346 )   PDF (1474KB) ( 175 )   Save BACKGROUND: Due to the multi-directional differentiation potential and other advantages, mesenchymal stem cells have become one of the most popular seed cells in tissue engineering. However, one of the major challenges is how to induce the cells into ideal targets. Mechanical stimulation plays a key role in directing the lineage commitment of mesenchymal stem cells. Plenty of studies exploring the mechanobiology of mesenchymal stem cells have been performed using in vitro loading to simulate the mechanical environment in the body. While progress has been made in understanding how mechanical signals are sensed by mesenchymal stem cells and then transduced to affect their behaviors (such as proliferation and differentiation), but the mechanotransductive mechanisms are still not fully understood. Stretch-activated channel is confirmed to play a significant role in the mechanotransduction of mesenchymal stem cells. OBJECTIVE: To summarize the studies on the mechanical regulation of mesenchymal stem cells, and then to analyze its role and possible mechanism in the mechanotransduction of mesenchymal stem cells. METHODS: We searched related articles in SCI and PubMed databases by “mesenchym al stem cell, stretch-activated channel, mechanical, tension, compression, fluid flow, hydrostatic pressure, piezo, TRP” as key words. Initially 160 articles related to mesenchymal stem cells and stretch-activated channel published from inception to April 2018 were searched, and 70 eligible articles were included in final analysis.  RESULTS AND CONCLUSION: Mesenchymal stem cells are important seed cells for tissue engineering because of their self-renewal ability and multi-directional differentiation potential. Researching the factors and mechanisms of their proliferation and differentiation gives a better access to clinical application. Mechanical factors play an indispensable role. Stretch, compression, fluid flow and hydrostatic pressure are four main mechanical stimulations related to the fate of mesenchymal stem cells. Stretch-activated channel forms a special group of mechanosensors that can serve as both sensors and effectors as they modify the electrical potential of the cell and mediate a flux of specific ions, such as Ca2+, across the plasma membrane. Then, these second messengers can associate with other ion channels, cytoskeleton, transcription factors to complete mechanotransduction.   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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Mechanism and application of stem cell transplantation in the treatment of ischemic stroke Zhang Dong-lin, Li Dan, Wang Min-juan 2018, 22 (33):  5393-5398.  doi: 10.3969/j.issn.2095-4344.0655 Abstract ( 295 )   PDF (671KB) ( 170 )   Save BACKGROUND: Once a nerve injury is developed after the onset of ischemic stroke, there is no effective treatment to promote the recovery of nerve function. A large number of experimental studies have proved that stem cell transplantation not only has neuroprotective effects, but also has the potential of cell replacement and functional recovery. OBJECTIVE: To review the specificity of stem cells, the possible mechanisms of stem cell transplantation for the treatment of ischemic stroke, the viable types of stem cells, the time, location and dose of stem cell transplantation, and the safety of surgery. METHODS: Using the keywords of “stem cells; transplantation; ischemic stroke” in English and Chinese, respectively, the first author retrieved relevant articles about stem cell transplantation in the treatment of ischemic stroke published from January 2013 to February 2018 in PubMed and CNKI databases. The literature types included clinical research, basic research and review articles. After removal of the articles that were not related to the purpose of the study or repetitive, 55 articles were finally analyzed. RESULTS AND CONCLUSION: With the development of technology and medicine, more types of stem cells have been discovered. However, due to the problems of cell origin, isolation and purification, and safety, mesenchymal stem cell transplantation for the treatment of ischemic stroke is the most widely used and studied. When selecting the timing, route, and dose of transplantation, 8×105 to 8×106 cells that are transplanted intravenously at 24 to 72 hours after ischemic injury have been widely used in basic or clinical experiments, which provide ideas for future research. However, whether it is the best method requires further experimental verification. Stem cell transplantation has broad application prospects in the treatment of ischemic stroke, and the value of stem cells has been repeatedly confirmed, which undoubtedly provides a new approach for the treatment of cerebral ischemia. However, this treatment method will encounter many challenges and there are still a lot of problems that need to be solved before it is used in clinical practice.  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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Treatment of complex anal fistula with adipose-derived stem cells: roles and mechanisms Liu Yan-ni, Ni Min, Zhang Rui, Huang Xiao-bo, Zhou Chun-gen, Jiang Bin 2018, 22 (33):  5399-5407.  doi: 10.3969/j.issn.2095-4344.0647 Abstract ( 369 )   PDF (959KB) ( 499 )   Save BACKGROUND: With the development of stem cell transplantation and tissue engineering technology, adipose-derived stem cell transplantation for complex anal fistula has received increasing attentions. By secreting various growth factors and cytokines, adipose-derived, stem cells regulate the body’s immune system, inhibit local and systemic inflammatory reactions, promote local angiogenesis, and activate fibroblasts, thereby accelerating wound healing. OBJECTIVE: To review the effects and existing problems of adipose-derived stem cells in the treatment of complex anal fistula as well as the mechanisms underlying wound healing. METHODS: The keywords of “adipose-derived stem cells, perianal fistulas, complex perianal fistulas, wound healing” in English and Chinese were to search relevant articles addressing adipose-derived stem cells for complex anal fistula treatment published from 2003 to 2018 in PubMed, Medline, CNKI, and WanFang. After removal of repetitive or irrelevant articles, 95 eligible articles were finally reviewed. RESULTS AND CONCLUSION: Adipose-derived stem cells accelerate postoperative wound healing by inducing differentiation, regulating inflammatory immunity, promoting angiogenesis, and activating fibroblasts. Clinical evidence from foreign literatures reveal that either autologous or allogeneic adipose-derived stem cells have special advantages in the treatment of anal fistula, including small trauma, no sphincter injury, little pain, rapid repair, low recurrence rate, and short hospital stay. Safety and effectiveness of adipose-derived stem cells for anal fistula has been initially confirmed, providing a new strategy for the clinical treatment of complex anal fistula. 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 inside and outside of the lung for lung repair: roles, functions and existing problems Chen Yan, Liu Xian 2018, 22 (33):  5408-5412.  doi: 10.3969/j.issn.2095-4344.0675 Abstract ( 321 )   PDF (546KB) ( 121 )   Save BACKGROUND: Stem cells have the ability to repair local tissue damage, which opens the possibility of cell therapy for most irreversible diseases, including chronic lung disease. OBJECTIVE: To elucidate the effect of stem cells in the repair of lung tissue. METHODS: The first author used the computer to retrieve CNKI and Medline databases using the keywords of “lung stem cells, tracheal and bronchial epithelial stem cells; transplantation; stem cells in the bronchial and terminal bronchial epithelium; stem cells at the bronchoalveolar junction; alveolar epithelial stem cells, interstitial lung stem cells” in Chinese and English, respectively. Approximately 500 articles were initially retrieved, and finally 47 eligible articles were included. RESULTS AND CONCLUSION: Stem cells inside and outside of the lung are the basis of lung injury, repair and remodeling. The lung stem cells in the body are mostly in a resting state. When the lung tissue is damaged or cultured in vitro, the stem cells from the lung or other tissues divide and proliferate. In addition, it is still unknown whether apoptosis, migration and differentiation will be triggered by other factors during the repair of lung tissue or even under physiological conditions. Therefore, many problems regarding stem cells for chronic lung diseases remain to be solved.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics