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08 September 2017, Volume 21 Issue 25 Previous Issue    Next Issue

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Effects of fibroblast growth factor on proliferation and differentiation of serially passaged bone marrow mesenchymal stem cells Song Ming-yu, Yang Yong, Wu Hua, Wang Rong 2017, 21 (25):  3937-3942.  doi: 10.3969/j.issn.2095-4344.2017.25.001 Abstract ( 345 )   PDF (1175KB) ( 192 )   Save BACKGROUND: The source of bone marrow mesenchymal stem cells (BMSCs) is limited, and the cellular morphology, proliferation and multi-directional differentiation capacities can vary during serial passages in BMSCs in vitro. OBJECTIVE: To study the effects of fibroblast growth factor (FGF) on cellular morphology, proliferation and differentiation of serially passaged BMSCs. METHODS: (1) BMSCs were isolated from Sprague-Dawley rats and cultured. These cells were passaged six times in vitro, and the cellular morphology was observed and photographed. (2) BMSCs at passage 6 were seeded into 96-well plates and randomly divided into control group and FGF treatment group. The proliferation of cells in both groups was detected with cell counting kit-8 kit at days 1, 2, 3, 4, 5, 6, 7 after culture. (3) BMSCs at passage 6 were seeded into 6-well plates and randomly divided into control group and FGF treatment group. After 7 days treatment with growth medium or growth medium containing FGF, the cellular morphology was observed and photographed. And then the cells of both groups were treated with osteogenic induction medium, adipogenic induction medium and chondrogenic induction medium for the next 7 days. The osteogenic, adipogenic and chondrogenic related genes (RUNX2, ALP, OCN; PPARγ2, AP2, ADIPOQ; SOX9, collagen II, aggrecan) were detected with real-time PCR. The protein expressions of RUNX2, PPARγ2, SOX9 were detected with western blot assay. (4) BMSCs at passage 6 were seeded into 6-well plates and randomly divided into control group and FGF treatment group. After 7 days treatment with growth medium or growth medium containing FGF, the cells were cultured with osteogenic induction medium, adipogenic induction medium and chondrogenic induction medium for the next 14 days. Then, alizarin red S staining, oil red O staining and alcian blue staining were performed. RESULTS AND CONCLUSION: (1) After in vitro passage for six times, the cellular morphology changed obviously, and FGF treatment recovered the characteristics of primary cells. (2) Compared with the control group, the cell proliferation in the FGF treatment group was significantly increased (P < 0.05). (3) Compared with the control group, the expression of osteogenic, adipogenic and chondrogenic related genes (RUNX2, ALP, OCN; PPARγ2, AP2, ADIPOQ; SOX9, collagen II, aggrecan) was increased significantly in the FGF treatment group (P < 0.05). The protein expressions of RUNX2, PPARγ2, SOX9 were also higher in the FGF treatment group than the control group (P < 0.05). (4) Compared with the control group, the number of extracellular calcium nodules, the number of intracellular lipid droplets, and the expression of acid acidic mucopolysaccharide were significantly increased after FGF pretreatment. To conclude, FGF pretreatment can preserve the stemness of BMSCs serially passaged in vitro.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Human umbilical cord Wharton’s jelly-derived mesenchymal stem cells protect against interleukin-1beta induced degeneration of nucleus pulposus cells Li Hao-yang, Chen Xu, Li Qiang, Ma Xun 2017, 21 (25):  3943-3948.  doi: 10.3969/j.issn.2095-4344.2017.25.002 Abstract ( 288 )   PDF (1115KB) ( 170 )   Save BACKGROUND: Human umbilical cord Wharton’s jelly-derived mesenchymal stem cells (hWJCs) via transplantation are able to survive in a dog model of intervertebral disc degeneration and to partially restore the height of the intervertebral disc. However, there is no study on whether hWJCs can repair nucleus pulposus cells after inflammatory injury as yet.  OBJECTIVE: To observe the protective effect of hWJCs on interleukin-1β induced rat nucleus pulposus degeneration. METHODS: Nucleus pulposus cells at passage 3 from Sprague-Dawley rats were divided into three groups: (1) normal control group cultured in culture medium containing fetal bovine serum for 24 hours, and then cultured in complete medium for 24 hours; (2) intervention group with interleukin-1β intervention for 24 hours, followed by culture in the complete medium for 24 hours; (3) co-culture group with interleukin-1β intervention for 24 hours, followed by culture in the complete medium for non-direct co-culture with hWJCs for 24 hours. Annexin V-FITC/propidium iodide (PI) double staining and flow cytometry were used in succession to measure the apoptosis rate of nucleus pulposus cells in each group. The ADAMTS-4, MMPs-3 and TIMP-1 expression in nucleus pulposus cells was observed though RT-PCR. The activities of caspase-3, caspase-8 and caspase-9 were determined by caspase kit. RESULTS AND CONCLUSION: The apoptosis rates were ranked as follows: the intervention group > co-culture group > normal control group, and there were significant differences between groups (P < 0.05). Compared with the control group, the expression levels of ADAMTS-4 and MMPs-3 were significantly higher and the expression level of TIMP-1 was significantly lower in the intervention and co-culture groups (P < 0.05). Compared with the intervention group, the expression levels of ADAMTS-4 and MMPs-3 were significantly lower and the expression level of TIMP-1 was significantly higher in the co-culture group (P < 0.05). The expression activities of caspase-3, caspase-8 and caspase-9 were highest in the intervention group, followed by the co-culture group, and lowest in the normal control group, and there were significant differences between groups (P < 0.05). To conclude, hWJCs has obviously inhibitory effect on the apoptosis of degenerated nucleus pulposus cells induced by interleukin-1β.   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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Genome-wide transcriptional profiling of NB4 leukemic cells affected by umbilical cord-derived mesenchymal stem cells Fan Hui-fang, Chen Fang, Ma Feng-xia, Chi Ying, Lu Shi-hong, Han Zhong-chao 2017, 21 (25):  3949-3955.  doi: 10.3969/j.issn.2095-4344.2017.25.003 Abstract ( 280 )   PDF (4939KB) ( 236 )   Save BACKGROUND: Mesenchymal stem cells (MSCs) are an important component of the in vivo microenvironment and act on multiple biological behaviors of tumor cells. The potential clinical value of MSCs has become an issue of concern in recent years. OBJECTIVE: To investigate the gene expression profiles of acute promyelocytic leukemia (APL) cell line NB4 treated with umbilical cord-derived MSCs (UC-MSCs) using cDNA microarray. METHODS: In vitro co-culture system was constructed, and then cellular proliferation, apoptosis and differentiation status of NB4 cells treated with UC-MSCs were evaluated. Two cDNA probes were prepared through reverse transcription from mRNA of NB4 cells treated with or without UC-MSCs. The probes were labeled with fluorescence dyes individually, hybridized with cDNA microarray, and their fluorescent intensities were scanned. The genes were screened through the analysis of the difference in two gene expression profiles. RESULTS AND CONCLUSION: UC-MSCs promoted the proliferation and differentiation, while reduced the apoptosis of NB4 cells. The analysis of gene expression profiles indicated that after co-culture with UC-MSCs, 530 genes were up-regulated and 53 genes were down-regulated. Accordingly, specific gene function and pathway signaling related were also regulated to some extent. Overall, UC-MSCs influence can major biological behaviors of NB4 cells by changing expression of a large amount of genes, gene-related function and multiple intracellular signaling pathways.    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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Mutant hypoxia inducible factor 1alpha transfection promotes proliferation of bone marrow mesenchymal stem cells Zhang Wei-wei, Wang Jue 2017, 21 (25):  3956-3963.  doi: 10.3969/j.issn.2095-4344.2017.25.004 Abstract ( 253 )   PDF (6912KB) ( 191 )   Save BACKGROUND: It has been found that hypoxia inducible 1α (HIF1α) gene can improve the ability of tissues and cells to survive in the ischemic environment, which is of great significance in promoting blood vessel regeneration. OBJECTIVE: To investigate the effect of the recombinant adenovirus Ad-CMV-HIF1αmu-IRES-hrGFP-1 on the proliferation of bone marrow mesenchymal stem cells. METHODS: Passage 3 bone marrow mesenchymal stem cells were cultured and transfected with Ad-CMV-HIF1αmu-IRES-hrGFP-1 virus (experimental group), with non-mutated gene Ad-CMV-HIF1α-IRES-hrGFP (positive control group), with Ad-CMV-IRES-hrGFP-1 empty virus (negative control group), or with no virus solution (blank control group). After transfection for 24 hours, the expression of HIF1α mRNA and protein was detected. Cell proliferation was detected by MTT assay at 24, 48, 72, 96 and 120 hours after transfection. RESULTS AND CONCLUSION: (1) Except that the mRNA expression of HIF1α in the experimental group and the positive control group was significantly higher than that in the negative control group and the blank control group (P < 0.05), there was no significant difference between two groups. (2) The expression of HIF1α protein in the experimental group was significantly higher than that in the positive control group, the negative control group and the blank control group (P < 0.05), and there was no significant difference between the latter three groups. (3) Cell proliferation was faster in the experimental group than the positive control group and the blank control group (P < 0.05), and moreover, there was no significant difference between the latter two groups. Our experimental findings indicate that the recombinant adenovirus-mediated three-point mutant HIF1α combined with hrGFP gene transfected bone marrow mesenchymal stem cells could not only express the target protein, HIF1α, under normoxic conditions, but also promote the proliferation of bone marrow mesenchymal stem cells.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of miR-218 and miR-26a on osteogenic differentiation of bone marrow mesenchymal stem cells by polyethyleneimine delivery Sun Xuan, Wei Ze-quan, Bu Wen-huan, Fang Tengjiaozi, Xiluopianduo, Liu Qi-lin, Sun Hong-chen 2017, 21 (25):  3964-3970.  doi: 10.3969/j.issn.2095-4344.2017.25.005 Abstract ( 329 )   PDF (7788KB) ( 185 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are widely used in the field of tissue engineering because of their multi-directional differentiation potential. Micro RNAs play an important role in promoting osteogenic differentiation of BMSCs. OBJECTIVE: To investigate the effect of miR-218 and miR-26a on the osteogenic differentiation of rat BMSCs, and to provide reference for the study on osteogenic differentiation of BMSCs and the clinical application. METHODS: The bone marrow of the femur of Wistar rats was extracted and the BMSCs were isolated and cultured to the 3rd generation. MiR-218, miR-26a and polyethyleneimine (PEI) were mixed in a specific ratio to form a miRNA/PEI complex. Meanwhile, a negative control group was established. RESULTS AND CONCLUSION: (1) Rat BMSCs grew well. (2) The optimal concentration of miRNA mimics was 50 nmol/L by MTT method. (3) The expression of alkaline phosphatase and collagen type I mRNA was significantly increased (P < 0.05). (4) Alkaline phosphatase staining showed that compared with the blank control group and the negative control group, the cytoplasm showed obvious coloring. (5) There were a lot of mineralized nodules shown by alizarin red staining. Therefore, miR-218/PEI complex, miR-26a/PEI complex and miR-218+miR-26a/PEI co-transfection complex could promote the osteogenic differentiation of BMSCs. Under the same conditions, the osteogenesis of miR-26a was slightly better than that of miR-218.   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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Age effects on in vitro differentiation and cytokine levels of macaque bone marrow mesenchymal stem cells Chen Yu-hao, Zhu Xiang-qing, Guo De-bin, Lin Qing-keng, He Zhi-xu, Pan Xing-hua 2017, 21 (25):  3971-3976.  doi: 10.3969/j.issn.2095-4344.2017.25.006 Abstract ( 346 )   PDF (4633KB) ( 177 )   Save BACKGROUND: Mesenchymal stem cells attract extensive attention because of good biological characteristics and broad prospects, but the cells gradually show the characteristics of the aging with the increase of individual age or incubation time in vitro. Nonhuman primates have similar biological characteristics with human being, and have unique advantage in the animal model and disease treatment research. OBJECTIVE: To analyze the difference in proliferation and differentiation of bone marrow mesenchymal stem cells from macaques at different ages and to explore the effect of age on bone marrow mesenchymal stem cells and the possible mechanism. METHODS: Bone marrow samples from male macaques aged < 3 years and over 20 years were collected through bone marrow puncture, and divided into young group and elder group, with three macaques in each group. Then, bone marrow mesenchymal stem cells were isolated and cultured in vitro, and the morphological changes, proliferation and differentiation ability were observed. Age-related beta-galactosidase staining was performed, and protein microarray and ELISA were used to detect cytokine levels. RESULTS AND CONCLUSION: With age, the proliferation and differentiation of bone marrow mesenchymal stem cells from the elder macaques were reduced significantly, and the number of senescent cells increased significantly; the levels of interleukin-1b, interleukin-4, interleukin-6, tumor necrosis factor α and vascular endothelial growth factor were elevated obviously, the levels of heparin-binding basic fibroblast growth factor and placental growth factor were reduced. These findings indicate that the body's aging lead to the reduction in the proliferation, differentiation and cytokine secretion of bone marrow mesenchymal stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Growth-associated protein-43 gene modified adipose-derived mesenchymal stem cells protect against retinitis pigmentosa Gao Jun-yan, Zhang Yun, Li Ming, Liu Xue-min, Wu Zhi-bing, Cao Yan-fei 2017, 21 (25):  3977-3982.  doi: 10.3969/j.issn.2095-4344.2017.25.007 Abstract ( 252 )   PDF (4457KB) ( 177 )   Save BACKGROUND: Gene therapy and cell transplantation therapy, especially their combined use, have certain therapeutic effects on retinitis pigmentosa. However, little attention has been paid to the combination of gene therapy and cell transplantation in the treatment of retinitis pigmentosa. OBJECTIVE: To investigate the protective effect of adipose-derived mesenchymal stem cells (ADMSCs) modified by growth-associated protein-43 (GAP-43) gene on the retina of rats with retinitis pigmentosa. METHODS: ADMSCs from rats with retinitis pigmentosa were isolated and cultured, and then transfected with GAP-43 lentiviral vector. Sixty retinitis pigmentosa rats were randomized into experimental group (subretinal injection of GAP-43-transfected ADMSCs), control group (subretinal injection of ADMSCs) and sham operation group (PBS injection). The expressions of GAP-43 protein, Rho protein and GS protein in the retina of rats were determined by western blot. The outer nuclear layer thickness of the retina was determined by hematoxylin-eosin staining. RESULTS AND CONCLUSION: After transfected cell transplantation, the expression of GAP-43 protein in the retina was gradually increased with time and showed significant differences at different time post-transplantation (P < 0.05). The expression levels of Rho protein and GS protein in the retina of experimental and control groups were higher than that in the sham operation group (P < 0.05). The expression of Rho protein in the retina of the experimental group was higher than that in the control group (P < 0.05). The expression of GS protein in the retina of the experimental group was lower than that in the control group (P < 0.05). The thickness of retinal outer nuclear layer was ranked as follows: the experimental group > the control group > the sham group, and there was significant difference between groups (P < 0.05). These results show that the ADMSCs modified by GAP-43 gene have protective effect on the retina of rats with retinitis pigmentosa.   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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Adipose-derived mesenchymal stem cells from hepatitis B virus infection patients differentiate into hepatocyte-like cells in vitro Wang Yan-hui, Wang Zhi-ru, Sun Yan, Zhao Wen-jing, Yang Shu-yuan, Zhao Shu-li 2017, 21 (25):  3983-3988.  doi: 10.3969/j.issn.2095-4344.2017.25.008 Abstract ( 339 )   PDF (4433KB) ( 201 )   Save BACKGROUND: It has been reported that human adipose-derived mesenchymal stem cells can be induced to differentiate into hepatocyte-like cells with the function of albumin synthesis and urea secretion in vitro. OBJECTIVE: To investigate the potential of adipose-derived mesenchymal stem cells differentiating into hepatocyte-like cells in vitro. METHODS: Adipose-derived mesenchymal stem cells were isolated from the subcutaneous fat of hepatitis B virus infection patients by collagenase digestion and adherent method. Adipose-derived mesenchymal stem cells  were induced by three-phase induction method and observed morphologically. The expression levels of alpha-fetoprotein, albumin and cytokeratin 18 were detected by immunohistochemical staining and glycogen synthesis function was detected by glycogen staining method. RESULTS AND CONCLUSION: Most of adipose-derived mesenchymal stem cells induced by three-phase induction method were differentiated into hepatocyte-like cells with polygonal morphology. Immunohistochemistry staining results showed that hepatocyte-like cells expressed alpha-fetoprotein, albumin and cytokeratin 18, and the expression levels of albumin and cytokeratin 18 increased with the culture time. The induced cells had the function of glycogen synthesis and were positive for periodic acid Schiff staining. These results showed that the subcutaneous adipose-derived mesenchymal stem cells could be induced into functional hepatocyte-like cells in hepatitis B virus infection 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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Phenotypic analysis of common cancer stem cell markers and cell sorting in lung cancer cell line A549 Peng Zhuo, Dang Xiao-yan, Zhang Qiu-hong, Zhang Rui, Yang Shuan-ying 2017, 21 (25):  3989-3994.  doi: 10.3969/j.issn.2095-4344.2017.25.009 Abstract ( 554 )   PDF (4084KB) ( 168 )   Save BACKGROUND: It has been widely reported that the most types of cancer are probably originated from cancer stem cells (CSCs) which are subpopulations of tumor cells. Recent studies also suggested that lung cancer could arise from CSCs. However, the phenotypic characteristics of CSCs in lung cancer have not been precisely described.  OBJECTIVE: To systematically analyze the expression of the most common CSC markers in cell line A549.  METHODS: A549 cells were cultured for 1 week under the condition of conventional high glucose. After that, flow cytometry was used to assess the expression of putative stem cell markers, including CD133, CD24, CD44 and ABCG2. Cells were then sorted according to the expression of CD44 and CD24 markers by fluorescence-activated cell sorting (FACS) and characterized using their sphere-forming capacity in serum free medium supplemented with several growth factors. RESULTS AND CONCLUSION: A549 cells expressed the CSC markers CD44 and CD24 at 64.23% and 58.62%, whereas the expression of both ABCG2 and CD133 was around 0.9%. Double-positive CD44/133 populations were rare. CD44+/CD24+ and CD44+/CD24- subpopulations respectively exhibited 54.64% and 23.38% expression. CD44+/CD24+ and CD44+/CD24- subsets were sorted by FACS. Both isolated subpopulations formed spheres in the serum free medium supplemented with basic fibroblast growth factor and epidermal growth factor. However, there was no significant difference in the sphere formation efficiency among CD44+/CD24+ and CD44+/CD24- subsets as well as A549 cells. Our findings suggest that CD44 and CD24 cannot be considered as potential markers for isolating lung CSCs in cell line A549, and further investigation using in vivo assays is required. 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 specific interfering TACC3 gene expression on proliferation and apoptosis of CD133+CD44+ oral squamous cell carcinoma cells Duan Rui, Li Yong-sheng, Xia Yi-chao 2017, 21 (25):  3995-4000.  doi: 10.3969/j.issn.2095-4344.2017.25.010 Abstract ( 276 )   PDF (1347KB) ( 188 )   Save BACKGROUND: Studies have indicated that the abnormal expression of TACC3 is closely related to the occurrence and development of many kinds of tumors, and the expression of TACC3 is up-regulated in these tumors. Therefore, in vitro specific inhibition of TACC3 expression may become an important target for the treatment or intervention of tumor growth. OBJECTIVE: To investigate the mechanism by which TACC3 gene expression regulates cell proliferation and apoptosis in oral squamous cell carcinoma. METHODS: CD133+CD44+ oral squamous cell carcinoma cells were sorted from human oral squamous cell carcinoma cell line Cal-27 by immunomagnetic beads. In experimental group, the shRNA sequence of TACC3 was designed and synthesized, which was then trasnfected into CD133+CD44+ oral cancer stem cells by LipofectamineTM 2000. Empty vector-trasnfected (negative control) and untransfected cells were used as callsed. Forty-eight hours after the transfection, effects of TACC3 gene silencing on proliferation and apoptosis in vitro in CD133+CD44+ oral squamous cell carcinoma were detected by MTT, clone formation test, and TUNEL assay. Western blot assay was used to detect the effect of TACC3 gene silencing on Ki67, Bax and Bcl-2 protein expression in CD133+CD44+ oral squamous cell carcinoma. RESULTS AND CONCLUSION: (1) Cell proliferation. The proliferation rate and expression level of Ki67 were significantly lower in the experimental group than the negative control and untransfected groups (P < 0.05). (2) Clone formation. The clone formation ability in the experimental group was significantly lower than that in the negative control and untransfected groups (P < 0.05). (3) Cell apoptosis. TACC3 gene silencing caused an obvious decrease in Bcl-2 protein expression and a significant increase in Bax protein expression. These findings further confirmed that specific interference of TACC3 gene expression could inhibit the proliferation of CD133+CD44+ cells and promote the apoptosis. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Curative effect of three-dimensional porous core decompression combined with autologous bone marrow transplantation on phase I, II avascular necrosis of the femoral head Wang Sen, Tan Mei-yun, Liu Shao-feng, Wu Tian-hao, Fan Zhong-wei, Zhang Yue, Rao Yong 2017, 21 (25):  4001-4006.  doi: 10.3969/j.issn.2095-4344.2017.25.011 Abstract ( 284 )   PDF (1162KB) ( 167 )   Save BACKGROUND: Hip-preserving treatment is advocated in the treatment of early avascular necrosis of the femoral head. OBJECTIVE: To compare the clinical efficacy of simple core decompression and three-dimensional porous core decompression combined with autologous bone marrow transplantation on early avascular necrosis of the femoral head.  METHODS: Thirty patients (39 hips) with early avascular necrosis of the femoral head at phase I-II according to the staging criteria of the Association Research Circulation Osseuse (ARCO) who had been admitted to the First Affiliated Hospital of Luzhou Medical University between March 2011 and May 2016 were surgically treated and followed up. Seventeen patients (22 hips, trial group) were treated with three-dimensional porous core decompression combined with autologous bone marrow transplantation, including 10 cases of ARCO stage I and 12 cases of ARCO stage II; the other 13 patients (17 hips, control group) were treated with simple core decompression, including 7 cases of ARCO stage I and 10 cases of ARCO stage II. Harris scores, X-ray and magnetic resonance imaging were used to evaluate the surgical efficacy at 1, 3, 6, 9, 12, 18, 24, 30, 36 months after the operation.  RESULTS AND CONCLUSION: The incision healed primarily in all patients, and no infection occurred. The follow-up lasted for 1-3 years (average 2.5 years). At the last follow-up, the Harris scores in both trial group and control group were significantly improved compared with the baseline (P < 0.05); moreover, the Harris scores of ARCO stage I and II patients were significantly higher in the trial group than the control group (P < 0.05). Collapse of the femoral head was observed in one hip (stage I) and one hip (stage II) of the trial group, and in three hips (stage I) and five hips (stage II) of the control group, and then hip arthroplasty was performed. The number of the patients developing collapse of the femoral head in the trial group was significantly less than that in the control group (P < 0.05). Compared with simple core decompression, three-dimensional porous core decompression combined with autologous bone marrow transplantation is more adept to improve patient’s function and delay disease progression in the treatment of ARCO stage I and II early avascular necrosis of the femoral head. 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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Ultrasound-mediated microbubble promotes bone marrow mesenchymal stem cell homing in the treatment of ischemic stroke Qian Jian, Fan Guo-feng, Xu Peng, Li Qi-ming, Sha Du-juan, Wang Jun, Zhang Jun 2017, 21 (25):  4007-4012.  doi: 10.3969/j.issn.2095-4344.2017.25.012 Abstract ( 388 )   PDF (1204KB) ( 173 )   Save BACKGROUND: In animal experiments, ultrasound-mediated microbubbles can promote the homing of transplanted stem cells to the ischemic area, enhance angiogenesis and small arterial formation, improve local blood flow in the ischemic myocardium and restore myocardial contractility. OBJECTIVE: To investigate the effect of ultrasound-mediated microbubbles on intravenously transplanted bone marrow mesenchymal stem cell (BMSC) homing and the therapeutic efficiency on ischemic stroke.  METHODS: A middle cerebral artery occlusion (MCAO) model was induced by plug wire preparation. At 72 hours after MCAO, model rats were randomized into four groups: PBS group (n=15), BMSCs group (n=18), ultrasound+BMSCs group (n=18), ultrasound+microbubble+BMSCs group (n=18). Corresponding treatment was done in each group: 2 mL of PBS was injected via tail vein in the PBS group; about 3×106 BMSCs diluted by 2 mL of PBS were injected via tail vein slowly in the BMSCs group; after skull ultrasound radiation (1 MHz, 2 W/cm2) for 120 seconds, BMSCs were injected via tail vein slowly in the ultrasound+BMSCs group; the same process as the ultrasound+BMSCs group was done following intravenous injection of 0.1 mL/kg microbubbles in the ultrasound+microbubble+BMSCs group. RESULTS AND CONCLUSION: (1) Forty-eight hours after BMSCs transplantation, the BMSCs homing rate in the brain was significantly higher in the ultrasound+microbubble+BMSCs group than the other two groups (P < 0.05). (2) Twenty-eight days after MCAO, nerve damage was significantly milder in the ultrasound+microbubble+BMSCs group than the other two groups (P < 0.05). (3) Seven days after transplantation, the water content in the brain tissue was significantly lower in the ultrasound+microbubble+BMSCs group than the other two groups (P < 0.05). (4) Seven days after transplantation, the cerebral infarction volume was significantly reduced in the ultrasound+microbubble+BMSCs group compared with the other two groups (P < 0.05). To conclude, ultrasound-mediated microbubbles can enhance the homing effect of intravenously transplanted BMSCs, reduce cerebral edema and cerebral infarction volume, improve the neurological function, and increase the therapeutic effect of BMSCs transplantation on ischemic stroke. 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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One-stage treatment with autologous red bone marrow transplantation for femoral comminuted fracture Pan Lin-lin, Wu Hao, Liu Guang-hui, Zhao Zhen-qun 2017, 21 (25):  4013-4019.  doi: 10.3969/j.issn.2095-4344.2017.25.013 Abstract ( 284 )   PDF (5619KB) ( 167 )   Save BACKGROUND: Whether one-stage bone healing with red bone marrow or mesenchymal stem cell transplantation is needed? Whether closed reduction with intramedullary interlocking nailing combined with autologous red bone marrow transplantation can promote osteogenesis in the one-stage treatment of femoral comminuted fracture still remains unclear. OBJECTIVE: To explore the effects of closed reduction with intramedullary interlocking nailing and one-stage treatment with autologous red bone marrow transplantation on fracture healing. METHODS: Twenty healthy New Zealand rabbits were selected and anesthetized, and then the models of femoral comminuted fracture were established at the hinder limbs after throwing a 5 kg of hammer from 30 cm height. All rabbits underwent intramedullary fixation using Kirschner wire (diameter: 2 mm); the right limbs served as control groups, and the left limbs as transplantation group, subjected to the injection of autologous red bone marrow (1 mL) into the fracture region. Then the rabbits were respectively killed at 14, and 28 days after modeling, the thickness of thickest callus was measured on X-ray films, and the number of chondrocytes and relative area of bone trabecula in the fracture region were observed through hematoxylin-eosin staining under light microscope. RESULTS AND CONCLUSION: (1) The thickness of callus formation in the control group was significantly less than that in the transplantation group at 14 days after modeling (P < 0.01). (2) The number of chondrocytes in the transplantation group was significantly more than that in the control group at 14 days after modeling (P < 0.01), and the fracture healing in the transplantation group was faster that in the control group. (3) Compared with the transplantation group, the relative area bone trabecula in the fracture region in the control group was significantly reduced at 28 days after modeling (P < 0.01). (4) These finding indicate that red bone marrow transplantation promotes fracture healing in the intramedullary fixation for femoral comminuted fracture in rabbits.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Combined use of low-dose 17-beta estradiol and bone marrow mesenchymal stem cell transplantation for spinal cord repair Kang Cong, Meng Xian-yong, Yang Xin-ming, Cheng Yao-yu, Zhang Zhen-liang 2017, 21 (25):  4020-4025.  doi: 10.3969/j.issn.2095-4344.2017.25.014 Abstract ( 293 )   PDF (5348KB) ( 180 )   Save BACKGROUND: Combined use of multiple interventions for different targets play superimposed or synergistic effects, which has become the current idea for spinal cord injury treatment. OBJECTIVE: To investigate the synergistic effects of low doses of 17-β estradiol combined with bone marrow mesenchymal stem cells (BMSCs) transplantation on the recovery of motor function and inflammatory reactions after spinal cord injury in rats.  METHODS: The 10 of 70 male Sprague-Dawley rats served as sham group in which the spinal cord was only exposed but with no treatment, and the rest 60 rats were used to make animal models of spinal cord injury using modified Allen’s method and then randomized into four groups (n=15 per group): model, estrogen, stem cell and combined treatment groups. Rats in the stem cell and combined treatment groups were given BMSCs transplantation at injured side; rats in the estrogen and combined treatment groups were given intramuscular injection of 17-β estradiol at 1 and 24 hours after modeling. At 1, 3, 5 and 7 days after modeling, rat functional recovery was evaluated by the Basso, Beatlie, Bresnahan score. The expressions of interleukin-1β and tumor necrosis factor-α in the injured spinal cord were detected by ELISA at 6, 12, 24, and 72 hours after modeling. Apoptosis in nerve cells was observed using TUNEL staining.  RESULTS AND CONCLUSION: The Basso, Beatlie, Bresnahan scores were declined significantly after modeling, increased at 5 and 7 days after stem cell transplantation, estrogen treatment or their combined treatment (P < 0.05), especially in the combined treatment group (P < 0.05). The levels of interleukin-1β and tumor necrosis factor-α were elevated gradually after spinal cord injury (P < 0.05), but the levels decreased significantly at 12 and 24 hours in stem cell, estrogen and combined treatment groups (P < 0.05), and this decrease trend was more significant in the combined treatment group compared with the stem cell and estrogen groups (P < 0.05). At 72 hours after modeling, the rate of TUNEL positive cells was highest in the model group (P < 0.05) and lowest in the combined treatment group (P < 0.05). To conclude, the combined use of low doses of 17-β estradiol and BMSCs transplantation can facilitate the recovery of motor function after spinal cord injury by effectively inhibiting apoptosis in nerve 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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Human umbilical cord mesenchymal stem cells via intraventricular injection improve the cardiac function of myocardial infarction rats Zhao Lu-yang, Sun Ying, Li Lian-chong 2017, 21 (25):  4026-4031.  doi: 10.3969/j.issn.2095-4344.2017.25.015 Abstract ( 313 )   PDF (5280KB) ( 149 )   Save BACKGROUND: Human umbilical cord mesenchymal stem cells (UC-MSCs), characterized by great differentiation potential, strong proliferation ability and low immunogenicity, exert an immeasurable role in wound repair and vascular regeneration in ischemic tissues. OBJECTIVE: To investigate the effect of intraventricularly injected UC-MSCs on expression of granulocyte-colony stimulating factor (GM-CSF) and transforming growth factor-β1 (TGF-β1) in the myocardium of myocardial infarction rats.  METHODS: 120 adult male Sprague-Dawley rats were selected randomly, among which, 20 healthy rats were randomly selected as controls and the rest rats were subcutaneously injected with isoproterenol to establish the myocardial infarction model. Model rats were randomized into model group, UC-MSC supernatant group, UC-MSC low-dose group, middle-dose group and high-dose group, with 20 rats in each group. Twenty-four hours after modeling, normal saline, UC-MSC supernatant, UC-MSC suspensions containing 0.25×106, 1.0×106, 4.0×106 cells (2 mL) were injected intraventricularly into the rats in the corresponding groups, respectively. Left ventricular ejection fraction (LVEF), end-systolic left ventricular volume (LVESV) and left ventricular end-diastolic volume (LVEDV) were measured by echocardiography at 2 weeks after treatment. Serum lactate dehydrogenase (LDH), creatine kinase (CK) and pro-brain natriuretic peptide (Pro-BNP) were measured by enzyme-linked immunosorbent assay (ELISA). At the end of the experiment, 10 rats were killed by dislocation and the cardiac specimens were taken. The myocardial infarct size was determined by nitroblue tetrazolium staining. The expressions of GM-CSF and TGF-β in the myocardium of rats in each group were determined by western blot method.  RESULTS AND CONCLUSION: (1) Higher LVEF levels and lower LVESV and LVEDV were found in the low-, middle- and high-dose UC-MSC groups than the model group and UC-MSC supernatant group (P < 0.05). LVEF, LVEDV and LVEDV in the middle- and high-dose UC-MSC group were lower than those in the low-dose UC-MSC group (P < 0.05). (2) The levels of serum LDH, CK, Pro-BNP and myocardial tissue GM-CSF and TGF-β1 in the low-, middle- and high-dose UC-MSC group were significantly lower than those in the model group and UC-MSC supernatant group (P < 0.05). Compared with the low-dose UC-MSC group, the levels of serum LDH, CK, Pro-BNP and myocardial tissue GM-CSF and TGF-β1 were significantly lower in the middle- and high-dose UC-MSC groups (P < 0.05). (3) The myocardial infarct sizes of the low-, middle- and high-dose UC-MSC groups were significantly lower than those of the model and UC-MSC supernatant groups (P < 0.05), while the myocardial infarct sizes of middle- and high-dose UC-MSC groups were significantly lower than that of low-dose UC-MSC group (P < 0.05). To conclude, UC-MSCs can significantly reduce the expression of GM-CSF and TGF-β1 in the rat myocardium after myocardial infarction, effectively protect myocardial tissues, and improve cardiac function. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Umbilical cord mesenchymal stem cells enhance imatinib-induced apoptosis in chronic myeloid leukemia Liu Ying, Song Bao-quan, Wei Yi-meng, Fan Hui-fang, Yu Yi, Dong Shu-xu, Han Zhong-chao, Ma Feng-xia 2017, 21 (25):  4032-4037.  doi: 10.3969/j.issn.2095-4344.2017.25.016 Abstract ( 343 )   PDF (4144KB) ( 182 )   Save BACKGROUND: Imatinib has a significant pro-apoptosis effect on chronic myelogenous leukemia (CML), but there are still some patients being resistant to it. Human umbilical cord mesenchymal stem cells (hUC-MSCs) affect the apoptosis of a variety of hematologic malignancies. However, the impacts of hUC-MSCs on the apoptosis of CML cells induced by imatinib remain unclear. OBJECTIVE: To investigate whether hUC-MSCs have an influence on the apoptosis of K562 cells induced by imatinib and to reveal the possible underlying mechanism. METHODS: K562 cells were cultured with hUC-MSCs or/and imatinib. Cellular apoptosis was measured with Annexin-V and PI staining by flow cytometry analysis. The protein expressions of Bax, Bcl-2, caspase-3, caspase-9 and cleaved-PARP in K562 cells were detected by western blot assay. Pan-caspase inhibitor Z-VAD-FMK was used to block apoptosis in each group, and during this process the effect of caspase apoptosis signaling pathway was detected. RESULTS AND CONCLUSION: The apoptosis of K562 cells was enhanced, when imatinib was combined with hUC-MSCs. Western blot analysis showed that the expression of pro-apoptotic protein Bax was enhenced and the expression of anti-apoptotic protein Bcl-2 was suppressed. Furthermore, the cleaved forms of caspase-9, caspase-3 and PARP in K562 cell were higher in the hUC-MSCs+imatinib group than in the imatinib group. The apoptosis of K562 cells induced by the hUC-MSCs combined with imatinib was significantly inhibited by Z-VAD-FMK. In conclusion, these findings indicate that hUC-MSCs can enhance imatinib-induced apoptosis of K562 cells by activating caspase apoptosis 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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Long non-coding RNA DANCR enhances chondrogenic differentiation and proliferation of human synovium-derived mesenchymal stem cells   Yang Chao, Zhang Lei, Zhou Li-wu, Zhao Jian-ning 2017, 21 (25):  4038-4043.  doi: 10.3969/j.issn.2095-4344.2017.25.017 Abstract ( 230 )   PDF (4307KB) ( 200 )   Save BACKGROUND: A number of studies have shown that long non-coding RNA DANCR can play an important role in various pathophysiological processes through Wnt/β-catenin signaling pathway. OBJECTIVE: To explore the effect of long non-coding RNA DANCR on the proliferation and chondrogenesis of synovium-derived mesenchymal stem cells. METHODS: Passage 3 synovium-derived mesenchymal stem cells were obtained and transfected with pcDNA3.1-GP (control) and pcDNA3.1-GP(DANCR Homo) (experimental). Cell viability was estimated at 1, 2, 3, 4, 5, 6 and 7 days after DANCR transfection. The passage 3 cells were cultured in the chondriogenic medium for 14 days. And the chondrogenesis potential of cells was examined by toluidine blue staining. The chondrogenic-specific marker genes Aggrecan, Type II collagen (Col2) and Sox9 were determined by Real-time PCR. RESULTS AND CONCLUSION: The synovium-derived mesenchymal stem cells exhibited “S”-shaped curves in the two groups, with cell arrest at 1-2 days and rapid proliferation beginning at 3 days. Cell counting kit-8 assay and toluidine blue staining showed overexpressing DANCR significantly promoted proliferation in synovium-derived mesenchymal stem cells. The aggregates from synovium-derived mesenchymal stem cells in the experimental group had a greater amount of toluidine blue staining than the control group. In addition, we detected the higher expression of chondrogenic specific marker genes, such as Col2, Sox9 and Aggrecan, in the experimental group than the control group at 14 days after chondrogenic induction (P < 0.05). These results demonstrate that long non-coding RNA DANCR could enhance chondrogenic differentiation and proliferation of human synovium-derived mesenchymal stem cells and increase the expression of chondrogenic specific marker genes. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Different types of allogeneic hematopoietic stem cell transplantation for Philadelphia chromosome-positive acute lymphoblastic leukemia: therapeutic efficacy and safety Hu Xiao-hui, Zhou Jie, Zhou Hai-xia, Zhang Ri, Wu De-pei 2017, 21 (25):  4044-4049.  doi: 10.3969/j.issn.2095-4344.2017.25.018 Abstract ( 370 )   PDF (1089KB) ( 182 )   Save BACKGROUND: With the optimization of transplantation scheme and the emergence of graft-versus-host disease (GVHD) therapy drugs, allogeneic hematopoietic stem cell transplantation (allo-HSCT) in recent years has made great progress that makes patients with hematological malignancies have more long-term survival opportunities. OBJECTIVE: To compare the efficiency and safety of three types of allo-HSCT used in the treatment of adults with Philadelphia chromosome (Ph) in acute lymphoblastic leukemia (Ph+ALL). METHODS: A total of 69 patients with Ph+ALL who received allo-HSCT from June 2006 to November 2013 were enrolled, including 23 cases of sib-matched donor transplantation, 24 cases of unrelated-matched donor transplantation, and 22 cases of haploidentical donor transplantation. There were 54 cases of CR1, 13 cases of CR2 to CR3 and 3 cases of relapse. The bone marrow or/and peripheral blood stem cells were used for transplantation. All patients were subjected to pretreatment consisting of cytarabine, busulfan, cyclophosphamide and total body irradiation. GVHD was prevented by combined use of immunosuppressants including cyclosporine A, short-term methotrexate, mycophenolate mofetil and anti-human thymocyte globulin, etc. RESULTS AND CONCLUSION: The results showed that 68 patients acquired hematopoietic reconstitution, and only 1 case of haploidentical donor transplantation failed. The mean follow-up period was 20.4 months. The acute GVHD incidence of the sibling matched-HSCT, unrelated donor HSCT and related haploidentical allo-HSCT was 30%, 33% and 45%, respectively; the chronic GVHD incidence (cGVHD) incidence was 22%, 29% and 36%, respectively; the incidence of aGVHD and cGVHD between groups showed no statistically significant difference. Transplant related mortality (TRM) was 9%, 29% and 41%, respectively, and there was a significant difference among groups (0.01 < P < 0.05). Recurrence rates were 17%, 21% and 14%, respectively, and there was no significant difference among groups. The 3-year overall survival rates were 68%, 49% and 43%, respectively; there were significant differences between sib-matched-HSCT and the other two groups, but no statistically significant difference was found between unrelated donor HSCT and related haploidentical allo-HSCT groups. The 3-year overall survival rate was 58% for 54 patients in CR1 and 41% for 15 patients in non-CR1 states. To conclude, the sib-matched HSCT has better effect than unrelated donor transplantation and related haploidentical allo-HSCT; Ph+ALL patients should do transplantation as early as possible in the state of CR1. 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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Role of sufentanil in neural stem cells transplantation for spinal cord injury Yan Hou-yong, Song Leng-mei, Liu Yue, Yao Guang, Zhang Ru-yi 2017, 21 (25):  4050-4056.  doi: 10.3969/j.issn.2095-4344.2017.25.019 Abstract ( 328 )   PDF (1181KB) ( 186 )   Save BACKGROUND: Studies have shown that the combined use of sufentanil and neural stem cell (NSC) transplantation can increase the number of newborn nerve fibers. OBJECTIVE: To investigate the effect of sufentanil on the hind limb function of rats with spinal cord injury after neural stem cell transplantation.  METHODS: (1) Eighty adult female Sprague-Dawley rats were used to build spine cord injury model according to the modified Allen’s method and divided into model group, sufentanil group, NSCs transplantation group and sufentanil combined with NSCs transplantation group (combined group). Extra 20 adult female Sprague-Dawley rats were not conducted any treatment as normal control group. (2) After 6 days of modeling, the model rats were subjected to subarachnoid injection of 10 μL of NSC culture medium and intraperitoneal injection of 100 μL of saline in the model group; subarachnoid injection of 10 μL of NSC culture medium and intraperitoneal injection of 100 μL of sufentanil(150 μg/kg) in the sufentanil group; subarachnoid injection of 10 μL of 1×1010/L NSCs suspension and intraperitoneal injection of 100 μL of saline in the NSCs transplantation group; and subarachnoid injection of 10 μL of 1×1010/L NSCs suspension and intraperitoneal injection of 100 μL of sufentanil (150 μg/kg) in the combined group. (3) After 72 hours of modeling, the AQP4 and MMP9 gene expression levels were detected by RT-PCR, and the cell apoptosis changes around the spine cord injury area were determined with TUNEL staining method. (4) The motor functions of rats were tested by Basso, Beattie and Bresnahan score and inclined plane test after 1, 3 days and 1, 2, 3 and 4 weeks of modeling. (5) After 4 weeks of modeling, the histopatholgical changes in the area of spine cord injury were observed by hematoxylin-eosin staining method. The survival changes of NSCs labeled by CM-Dil were determined by fluorescence microscope. The regenerations and distributions of spinal nerve fibers were observed by fluorescein gold retrograde tagging.  RESULTS AND CONCLUSION: (1) After 72 hours of modeling, the AQP4 and MMP9 gene expression levels as well as the cell apoptotic rate in the combined group were significantly lower than those in the model, sufentanil and NSCs transplantation groups (P < 0.05). (2) After 2 weeks of modeling, the combined treatment significantly improved the hind limb motor functions of rats compared with the sufentanil and NSCs transplantation groups (P < 0.05), and the recovery of motor function was better in the sufentanil and NSCs transplantation groups than in the model group (P < 0.05). (3) After 4 weeks of modeling, the results of hematoxylin-eosin staining manifested that the spinal cord tissues lost and the magnified syringomyelias occurred in the model group. The syringomyelias in the sufentanil and NSCs transplantation groups were significantly smaller than that in the model group, and the syringomyelias almost disappeared in the combined group. (4) The number of positive cells was the most in the combined group, more in the NSCs transplantation group, but there were no positive cells labeled by CM-Dil in the sufentanil and negative control groups. (5) The number of positive neural fibers in the combined group was the highest followed by the sufentanil and NSCs transplantation groups, and the number of positive neural fibers in negative control group was the lowest. To conclude, sufentanil can improve the recovery of hind limb motor function by reducing the AQP4 and MMP9 expression levels in the injury area, promoting the survival of transplanted NSCs, and decreasing the local NSCs apoptosis after spinal cord injury. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Induced pluripotent stem cells differentiate into intestinal organoids in three-dimensional niche in vitro Li Xiang-yang, Zhao Xin, Xiang Xiao-song, Zheng Peng, Hui Huang, Ji Wu 2017, 21 (25):  4057-4061.  doi: 10.3969/j.issn.2095-4344.2017.25.020 Abstract ( 293 )   PDF (1036KB) ( 191 )   Save BACKGROUND: Induced pluripotent stem cells (iPSCs) are a special type of cells with self-renewal and multi-differentiation potential, which can differentiate into intestinal organoids under certain conditions. OBJECTIVE: To explore whether iPSCs can differentiate into intestinal organoids under specific conditions in vitro. METHODS: iPSCs from B6J mice were recovered and cultured for 3 days until clone units covered about 80% of the culture dish, and then the cells were cultured in the medium containing Activin A for 3 days until the deterministic endoderm formed. Further, the culture medium was replaced by the medium with fibroblast growth factor 4 and Wnt3A for 4 days to differentiate into the spheroids with CDX2+. After that, spheroids were collected and mixed with Matrigel, and then the mixture was dropped into the 4-well plate and cultured with Rspondin1, Noggin, epidermal growth factor, B27 and other growth factors to differentiate into intestinal organoids. Cell morphology was observed, FoxA2 and Sox17 expresson in the deterministic endoderm was detected, and CDX2, Sox9, CGA, MMP7 were measured.  RESULTS AND CONCLUSION: iPSCs were cultured with Activin A for 3 days with higher cell fusion, initial differentiation and FoxA2/Sox17 expression (P < 0.05) than those of non-induced iPSCs. Spheroids began to appear at the 3rd day after culture with fibroblast growth factor 4 and WNT3A, and formed a lot at the 4th day. And CDX2 expression in spheroids was significantly increased compared with that in the deterministic endoderm (P < 0.05). Organoids gradually formed after 3 days culture, which contained all cell types of intestinal organoids, and expressions of specific markers, Sox9, CGA, MMP7, were significantly higher than those in spheroids (P < 0.05). To conclude, iPSCs can be induced to differentiate into intestinal organoids in three-dimensional niche in vitro. 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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Genetic polymorphism of 12 short tandem repeat on chromosome X loci in Shenzhen, China: a pedigree investigation Li Zhen, Li Xue-mei, Zou Hong-yan, Cheng Liang-hong 2017, 21 (25):  4062-4067.  doi: 10.3969/j.issn.2095-4344.2017.25.021 Abstract ( 278 )   PDF (1039KB) ( 170 )   Save BACKGROUND: The special genetic law of short tandem repeat on chromosome X (X-STR) makes it incomparable with autosome markers in forensic identification. However, the population genetics data is far less than that of the autosome STR, and especially the haplotype data are rarely reported. OBJECTIVE: To study the genetic polymorphism of 12 X-STR loci in Shenzhen area by pedigree analysis, aiming to provide scientific and effective data for the application of X-STR in forensic medicine and genetics. METHODS: The blood samples of 118 families were taken to extract DNA by Chelex-100, followed by PCR amplification using Investigator Argus X-12 kit. The frequency of alleles of 231 unrelated individuals was counted by direct counting method and Excel software. Hardy-Weinberg equilibrium test was performed on 12 X-STR loci of female samples by chi-square test. Discrimination power and mean exclusion chance were calculated according to the formula. Pedigree analysis was done to identify haplotypes of female samples and the haplotype frequencies of 4 linkage groups in 111 fathers and 119 mothers were calculated using direct counting method and Excel software. RESULTS AND CONCLUSION: In this study, 349 haplotypes were obtained. There were 238, 139, 153 and 157 haplotypes in linkage groups X1-X4, respectively. The polymorphism of DXS10135 locus was the highest with 21 alleles, while the polymorphism of DXS7423 locus was the worst with only 4 alleles. The combined discrimination power was 0.99999999 in males and 0.99999999 in females. The combined mean exclusion chance was 0.99999999 in trio cases, and 0.99999811 in duo cases. These findings indicate that the X-12 detection system has high polymorphism in Shenzhen Han population, and has important application value in forensic individual identification and paternity testing. 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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Identification of genomic full-length sequence of human leukocyte antigen-E and its two novel alleles He Liu-mei, Wang Song-xing, Xu Yun-ping 2017, 21 (25):  4068-4074.  doi: 10.3969/j.issn.2095-4344.2017.25.022 Abstract ( 319 )   PDF (2828KB) ( 209 )   Save BACKGROUND: Human leukocyte antigen-E (HLA-E) is one of non-classical HLA class I genes. Up to now, the polymorphism analysis is mainly aimed at the variation in exon 3 of HLA-E, which determines HLA-E*01:01 or HLA-E*01:03. However, the identification of the full-length HLA-E and its novel alleles is rare reported. OBJECTIVE: To establish the method of identification of HLA-E genomic full-length sequence, and to identify its novel alleles in healthy blood donors in Shenzhen, China. METHODS: Peripheral blood DNA samples were extracted from the subjects, and the amplified primers and sequencing primers in conserved regions were designed according to the sequences of HLA-E published in the IMGT/HLA database. A high-fidelity reaction system was used to amplify the genomic full-length of HLA-E, followed by sequencing, assembling, confirming and typing. RESULTS AND CONCLUSION: Herein, we successfully established the method for amplifying genomic full-length sequence and sequence-based typing. Two novel HLA-E alleles were nominated by WHO HLA Nomenclature committee as HLA-E*01:01:01:06 and HLA-E*01:01:01:07. Compared with the most related allele HLA-E*01:01:01:01, HLA-E*01:01:01:06 had one nucleotide change at nt-26(G->T) in 5’-promoter, and HLA-E*01:01:01:07 had one nucleotide change at nt3345(T->C) in 3’-UTR. The polymorphism data of genomic full-length HLA-E in Chinese individuals need to be filled, and the method we developed here supplies the key technique for the further studies.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Application of human acelluar amniotic membrane in tissue engineered scaffold construction Yang Ji-bin, Xiong Hua-zhang, Li Yu-wan, Liu Yi 2017, 21 (25):  4075-4081.  doi: 10.3969/j.issn.2095-4344.2017.25.023 Abstract ( 259 )   PDF (1128KB) ( 205 )   Save BACKGROUND: Human acellular amniotic membrane is a kind of extracellular matrix material with good biocompatibility and biological activity. It has been widely used in various clinical studies because of its low immunogenicity, small rejection and easy preparation. OBJECTIVE: To review the applications of human acellular amniotic membranes in tissue engineering field, such as skin, blood vessel, cornea, cartilage and bone. METHODS: CNKI (from January 2005 to May 2017), CBMdisc (from January 2005 to May 2017), PubMed (from January 1990 to May 2017) and Elsevier (from January 1990 to May 2017) were retrieved for articles addressing the application of human acellular amniotic membrane as a tissue-engineered scaffold in the bone, cartilage, skin, and blood vessels. The keywords were “acelluar amniotic membrane, scaffold, material, tissue engineering ECM" in Chinese and English, respectively. RESULTS AND CONCLUSION: Human acellular amniotic membrane owns the structure and function of the natural extracellular matrix, which can be combined with stem cells from different sources to differentiate into different tissues and organs, such as bone, cartilage, skin, blood vessel, and corneal tissues. As a tissue-engineered scaffold, human acellular amniotic membrane has good biocompatibility, biodegradability and low immunogenicity, although it has some shortcomings, such as poor strength and post-transplantation rejection reactions. Therefore, the future studies are mainly focused on shortening the adhesion time between cells and scaffolds, increasing the own mechanical strength of human acellular amniotic membrane, optimizing the cell growth microenvironment, and combining human acellular amniotic membrane with other tissue-engineered scaffolds.  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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Directional differentiation of bone marrow mesenchymal stem cells Li Qiao-qiao, Wu Zhen-qiang, Zhang Li-jun 2017, 21 (25):  4082-4087.  doi: 10.3969/j.issn.2095-4344.2017.25.024 Abstract ( 309 )   PDF (961KB) ( 191 )   Save BACKGROUND: Bone marrow mesenchymal stem cells is the most promising tissue engineering seed cells because of their strong proliferation and passage ability, multi-directional differentiation potential, as well as no ethical problems and no rejection. OBJECTIVE: To review the recent advances in the directional differentiation of bone marrow mesenchymal stem cells induced by various conditions METHODS: The first author retrieved CNKI, PubMed, Wanfang databases for relevant articles published from January 2001 to September 2016. The keywords were “bone marrow mesenchymal stem cells, BMSCs, directional differentiation” in Chinese and English, respectively. Literature addressing experimental research and review were selected, and finally 48 articles were included. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells can be induced in vitro to differentiate into chondrocytes, osteoblasts, adipocytes, and nerve cells by cytokines, chemical drugs, Chinese herbs and extracts, physical stimulation. To establish feasible induction conditions in vitro and improve cell induction efficiency is the basis for the clinical application of bone marrow mesenchymal stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Anti-inflammatory effect of stem cells in the treatment of ischemic stroke Gao Feng-juan, Gao Shan-e, Chen Xu, Sun Jian, Wang Jun-yi 2017, 21 (25):  4088-4093.  doi: 10.3969/j.issn.2095-4344.2017.25.025 Abstract ( 327 )   PDF (877KB) ( 226 )   Save BACKGROUND: A large number of preclinical experimental data have shown that stem cells can regulate the immune function, and serve the function of cell replacement. OBJECTIVE: To summarize the anti-inflammatory effects of stem cells in the treatment of ischemic stroke, based on which, we further discuss the specific mechanisms. METHODS: We conducted a systematic and comprehensive search in PubMed, Elsevier, Springer, Wiley, Ovid, EBSCO databases. The range of retrieval time was from 2012 to 2017. The keywords were “stem cells, stroke, inflammation, immune”. Totally 110 articles were retrieved initially, and 47 articles were included in result analysis. RESULTS AND CONCLUSION: By reading extensive literature, we analyzed and summarized the research status quo of the anti-inflammatory effects of neural stem cells, mesenchymal stem cells and endothelial cells in the treatment of ischemic stroke. The mechanisms mainly include reducing focal inflammation, immune regulation, promoting the secretion of various neurotrophic factors, reducing secondary cell death, protecting neurons and promoting cell function recovery and further promoting the recovery of nerve function. The mechanisms underlying local immune regulation and anti-inflammatory effects of stem cells are mainly described as the shift from M1 to M2 macrophages under the intervention of stem cell factors, to intervene secreted immune cytokine profiles and exert effects on inhibition and polarization of glial cells. Further investigation is required on the anti-inflammatory effects and immune regulation of stem cell therapy for stroke. 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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Current status of research on bone marrow necrosis syndrome Mao Hui, Jia Wen-hua, Huang Yi-hong, Li De-peng 2017, 21 (25):  4094-4100.  doi: 10.3969/j.issn.2095-4344.2017.25.026 Abstract ( 589 )   PDF (1078KB) ( 156 )   Save BACKGROUND: Bone marrow necrosis has unspecific clinical features, which is often misdiagnosed or missed due to a lack of the knowledge of the disease. OBJECTIVE: To improve the awareness and vigilance to bone marrow necrosis, and to further explore the clinical manifestations, hematological characteristics, pathogenesis and treatment of bone marrow necrosis. METHODS: The “bone marrow necrosis, hematologic neoplasms, solid tumor, bone marrow puncture, bone marrow pathology” in Chinese and English served as the search terms to search articles related to bone marrow necrosis in PubMed and Wanfang databases, published from 1941 to 2016. Totally 43 articles were selected for review. RESULTS AND CONCLUSION: Bone marrow necrosis is a rare complication caused by various diseases, clinically characterized by bone pain, fever, anemia, and nucleated red cells and immature neutrophilic leukocytes in the blood smear. Bone marrow aspiration and/or bone marrow biopsy show(s) necrotic features. Its pathogenesis is complex, and it is still poorly understood and needs further research. There is no good treatment for bone marrow necrosis, and the prognosis is poor. Early correct diagnosis and etiological treatment are crucial for the prognosis of bone marrow necrosis. With the improvement of disease awareness, bone marrow cytology, genetics, MRI and hematopoietic stem cell transplantation, bone marrow necrosis is expected to get a better prognosis. 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