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05 August 2016, Volume 20 Issue 32 Previous Issue    Next Issue

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Mechano growth factor promotes the differentiation of bone marrow mesenchymal stem cells into osteoblasts Tong Yan-xiang, Feng Wei, Jia Yan-fei, Wang Cai-xia, Lv Hui-cheng, Wu Yi-min, Jiang Dian-ming 2016, 20 (32):  4717-4724.  doi: 10.3969/j.issn.2095-4344.2016.32.001 Abstract ( 287 )   PDF (1290KB) ( 257 )   Save BACKGROUND: Mechano growth factor has the potential to activate muscle satellite cells and promote myogenic cell growth, and has dual roles in maintaining bone mass and repairing bone defects. OBJECTIVE: To explore the mechanism underlying osteogenic differentiation of rabbit bone marrow mesenchymal stem cells promoted by the mechano growth factor. METHODS: The best concentration and time of mechano growth factor to promote osteogenic differentiation of rabbit bone marrow mesenchymal stem cells were detected by MTT. The mRNA and protein expressions of alkaline phosphatase and osteocalcin were detected by qPCR and western blot, respectively. The phosphorylation level of AKT and mTOR were detected by western blot assay. RESULTS AND CONCLUSION: The best concentration and time of mechano growth factor was 45 μg/L and 5 days for promoting the osteogenic differentiation of rabbit bone marrow mesenchymal stem cells. The expressions of alkaline phosphatase and osteocalcin at mRNA and protein levels were highest after 4-hour intervention with 45 μg/L mechano growth factor, and meanwhile, the phosphorylation levels of mTOR and AKT were also highest. These findings indicate that the mechano growth factor can promote the differentiation of rabbit bone marrow mesenchymal stem cells into osteoblasts via PI3K/AKT pathway, and its best concentration and time are 45 μg/L and 4 hours, respectively.     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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Perfusion in vivo of bone marrow stromal stem cells to build tissue-engineered bone for treating benign bone tumors and tumor-like lesions Wang Shun-li, Shi Ying-bin, Zhang Lin-feng 2016, 20 (32):  4725-4730.  doi: 10.3969/j.issn.2095-4344.2016.32.002 Abstract ( 280 )   PDF (1154KB) ( 248 )   Save BACKGROUND: Bone marrow stromal stem cells have a strong osteogenic potential, which are currently the most ideal seed cells for tissue engineering. However, there is no clinical report on the treatment of benign bone tumors and tumor-like lesions using bone marrow stromal stem cell transplantation. OBJECTIVE: To investigate the in vivo perfusion method of inducing bone marrow stromal stem cells, and the clinical effects of bone marrow stromal stem cells on benign bone tumors and tumor-like lesions. METHODS: Sixty-five cases of benign bone tumors and tumor-like lesions were divided into three groups according to the different treatments: bone graft group (n=30) and bone marrow stromal stem cells group (n=35). In the bone graft group, allogeneic bone was soaked in normal saline for 30 minutes, and then implanted into the bone defect site. In the bone marrow stromal stem cells group, 20-40 mL of bone marrow from each patient was extracted to isolate, purify and culture bone marrow stromal stem cells that were then perfused into the bone defect site. RESULTS AND CONCLUSION: Under the inverted phase contrast microscope, the perfused cells appeared as a spherical shape, with different sizes. Initially, there were more hematopoietic cells in the perfusion cell culture. With the extension of the culture time, adherent spindle cells and suspended red blood cells appeared, which were mostly round and triangular. All the patients were followed up for 1-12 months and healed well after surgery. Compared with the bone graft group, infection rate and healing time were both lower in the bone marrow stromal cell group. To conclude, in vivo perfusion of bone marrow stromal stem cells used for construction of tissue-engineered bone promotes blood supply reconstruction and bone healing in patients with benign bone tumors and tumor-like lesions, which is of high clinical values.     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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Curcumin effect on proliferation and apoptosis of gastric cancer stem cells via ATK/FoxM1 signaling pathway He Dong-li 2016, 20 (32):  4731-4737.  doi: 10.3969/j.issn.2095-4344.2016.32.003 Abstract ( 363 )   PDF (1343KB) ( 296 )   Save BACKGROUND: Curcumin has crucial inhibitory effects on various cancer cells and cancer stem cells. However, its effect on gastric cancer stem cells and the underlying mechanism of this effect are unclear. OBJECTIVE: To explore the effect of curcumin on gastric cancer stem cells and the underlying mechanism. METHODS: Tumor sphere-forming assay and gastric cancer stem cell markers (EpCAM and CD44) were used to separate gastric cancer stem cells from gastric cancer SGC7901 cell lines. Effects of curcumin on the proliferation and apoptosis of gastric cancer stem cells were determined by MTT and flow cytometry analysis, respectively. Western blot analysis was used to detect the expression levels of FoxM1, p-AKT, and AKT. LY294002, an inhibitor of the PI3K/AKT pathway, was used to determine the regulatory relationship between AKT and FoxM1 signaling pathways. RESULTS AND CONCLUSION: The EpCAM+/CD44+ gastric cancer stem cells were successfully isolated from SGC7901 cells. MTT assay showed that curcumin inhibited the proliferation of gastric cancer stem cells, while flow cytometry analysis showed that curcumin induced apoptosis in gastric cancer stem cells. In addition, the expression levels of p-AKT and FoxM1 were decreased by curcumin treatment. After being treated by LY294002, the expression levels of p-AKT and FoxM1 were down-regulated markedly. In conclusion, curcumin can inhibit cell proliferation and induce apoptosis in gastric cancer stem cells via the ATK/FoxM1 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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Gastric cancer stem cells in tumor invasion and metastasis and its influence on angiogenesis ability Zhang Li-xian, Zhang Ning, Yuan Shuang-zhen, Chen Yu-mei, Su Xin-ai, Sun Jian-shun, Wang Li-hua, Lin Mei 2016, 20 (32):  4738-4744.  doi: 10.3969/j.issn.2095-4344.2016.32.004 Abstract ( 263 )   PDF (1143KB) ( 194 )   Save BACKGROUND: Gastric cancer stem cells involved in the neoadjuvant chemotherapy and conventional treatment are closely associated with relapse of gastric cancer. However, this conclusion has not yet been confirmed. OBJECTIVE: To investigate the effects of gastric cancer stem cells in tumor invasion and metastasis and its effect on angiogenesis ability. METHODS: We prepared nude mouse models of gastric cancer to isolate and culture gastric cancer stem cells. Harvested gastric cancer stem cells were detected in cell scratch test, ring test, inhibition rate test, cell migration test and tumorigenicity test. RESULTS AND CONCLUSION: After 7 days of culture, the cells exhibited adherent growth but a lack of regularity that most cells were in a tadpole shape. In the cell scratch test, the scratch width was significantly different at 0 and 24 hours (P < 0.05). Under an inverted microscope, the cells were found to form a ring in the ring test. The 50% inhibiting concentration of gastric cancer stem cells induced by oxaliplatin was significantly lower than that induced by 5-fluorouracil (P < 0.05). The number of cells passing through the basilar membrane was significantly increased after cell migration (P < 0.05). After implantation of gastric cancer stem cells, the gastric tissue quality was significantly higher than that in normal nude mice of gastric cancer (P < 0.05). These findings indicate that gastric cancer stem cells responsible for tumor invasion and migration have stronger angiogenesis ability.     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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Neuron-like differentiation of bone marrow mesenchymal stem cells from children in vitro Li Ying, Gao Jie, Shang Ya-min, Wang Qi-wei, Wang Hai-lei 2016, 20 (32):  4745-4750.  doi: 10.3969/j.issn.2095-4344.2016.32.005 Abstract ( 243 )   PDF (3461KB) ( 260 )   Save BACKGROUND: In the past, the culture and differentiation of bone marrow mesenchymal stem cells in vitro were mostly reported in the adult or animal rather than in children. OBJECTIVE: To explore the ability of bone marrow mesenchymal stem cells from children differentiating into neural stem cells and nerve cells. METHODS: Bone marrow mesenchymal stem cells from children were isolated and cultured, and passage 12 cells were cultured in the pre-induction medium (DMEM culture medium containing 10% fetal bovine serum and 1 mmol/L β-mercapto ethanol) and induction medium (DMEM containing 2% dimethyl sulfoxide and 150 μmol/L butylated hydroxyanisole). Expression of nestin and β-tublin III was detected using immunocytochemistry method at 30 minutes and 7 days after induction, while RT-PCR was used to detect nestin mRNA expression at 0, 5.5, 6 days after induction. RESULTS AND CONCLUSION: After combined induction, the cells shrank from round shape to tapered, polygonal or oval shape, and cell processes extended gradually and became filament-like shape. Interconnected cells formed a network at 6 days after combined induction. The expression of nestin antigen was positive at 30 minutes after induction, while the expression of β-tublin was positive at 7 days. RT-PCR findings showed that positive expression of nestin mRNA was detected at 5.5 hours of induction, and then disappeared at 6 days. These findings show that the combined use of dimethyl sulfoxide and butylated hydroxyanisole can induce bone marrow mesenchymal stem cells from children to differentiate into neural stem cells and nerve cells 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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Effects of endocrine therapy and chemotherapy on stem cells in patients with breast cancer Zhao Yan-li, Diao Jian-hua, Wu Cai-qin, Zhang Cai-xia, Jia Huan-xia, Niu Zhao-xia 2016, 20 (32):  4751-4757.  doi: 10.3969/j.issn.2095-4344.2016.32.006 Abstract ( 354 )   PDF (4995KB) ( 257 )   Save BACKGROUND: Tumor stem cells are the root of cancer recurrence and metastasis, so clinical researches should focus on the effects of different treatments on tumor stem cells. OBJECTIVE: To explore the effects of endocrine therapy and chemotherapy on stem cells in patients with breast cancer. METHODS: After recovery and cultivation of estrogen receptor-positive human breast cancer cell lines MCF-7, passage 3 cells in logarithmic phase were selected and divided into three groups containing control, estradiol and estradiol with tamoxifen groups. The estradiol group was divided into three subgroups: 10-7, 10-8 and 10-9 mol/L estradiol was added into the medium, respectively; the estradiol with tamoxifen group was divided into three subgroups: 10-7, 10-8 and 10-9 mol/L estradiol with 10-6 mol/L tamoxifen were added into the medium, respectively. The same amount of absolute ethyl ethanol was added into the medium of control group. Fifteen female patients with late recurrence and metastasis of breast cancer received chemotherapy as recurrence and metastasis group. Another 15 healthy volunteers were selected as healthy control group. RESULTS AND CONCLUSION: The proportion of CD44+CD24-/low cell subsets in the estradiol and estradiol with tamoxifen groups was significantly higher than that of the control group (P < 0.05), and the proportion of CD44+CD24-/low cell subsets in the estradiol group was significantly higher than that of the estradiol with tamoxifen group at the same concentration (P < 0.05). The proportion of CD44+CD24-/low cell subsets had no significant differences among groups at 10 and 20 days of culture (P < 0.05). The proportion of CD44+CD24-/low cell subsets significantly increased in MCF-7 cells after 24-hour intervention with different chemotherapy drugs. But only the proportion of CD44+CD24-/low cell subsets in the paclitaxel and doxorubicin groups was significantly higher than that of the control group after 20-day intervention (P < 0.05). Besides, the proportion of CD44+CD24-/low cell subsets in the peripheral blood of healthy volunteers was significantly lower than that of the recurrence and metastasis group (P < 0.05). Among 15 patients with late recurrence and metastatic of breast cancer, 9 had stable disease, 5 had partial remission, 1 had failed chemotherapy and cancer progression. Moreover, the proportion of CD45-CD44+CD24-/low cell subsets in the peripheral blood of patients sensitive for chemotherapy was significantly lower than that before treatment (P < 0.05). In conclusion, both endocrine therapy and chemotherapy exert a certain effect on the CD44+CD24-/low cell subsets of breast cancer positive for estrogen receptor. Given that CD44+CD24-/low cell subsets in MCF-7 cells resist chemotherapy drugs, the proportion of CD45-CD44+CD24-/low cells in the peripheral blood of patients sensitive for chemotherapy is decreased.     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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Relationship between cell activity and multidrug resistance of CD44+CD24-/low breast cancer stem cells Zhai Xiao-jian, Zhang Hao, Zhang Yi-ni, Ni Ming, Wang Zheng 2016, 20 (32):  4758-4763.  doi: 10.3969/j.issn.2095-4344.2016.32.007 Abstract ( 348 )   PDF (3623KB) ( 219 )   Save BACKGROUND: Tumor stem cells are found to be involved in the recurrence, metastasis and drug resistance of the tumor. OBJECTIVE: To explore the relationship between cell activity and multidrug resistance of CD44+CD24-/low breast cancer stem cells. METHODS: CD44+CD24-/low breast cancer stem cells sorted from multidrug resistant breast cancer cell line MCF-7/ADR were detected as percentage using flow cytometry. P-gp fluorescence intensity of the cell membrane and MDR mRNA expression in sorted cells and MCF-7/ADR were detected using flow cytometry and RT-PCR, respectively. RESULTS AND CONCLUSION: After sorting by flow cytometry, the proportion of CD44+CD24-/low breast cancer stem cells was more than 90%, indicating that the sorted cells could meet the needs of the subsequent experiment. CD44+CD24-/low cell subsets exhibited stronger ability to form microspheres than non- CD44+CD24-/low cell subsets. The P-gp fluorescence intensity and MDR mRNA expression of CD44+CD24-/low cells were significantly higher than those of MFC-7/ADR cell line (P < 0.05). These experimental findings suggest that CD44+CD24-/low breast cancer stem cells sorted from MCF-7/ADR cell lines have a strong ability to form cell microspheres in vitro, and significantly raise the level of P-gp protein and MDR mRNA expression, which may be one of the causes of multidrug resistance.     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 blood stem cells differentiate into nasal ciliated epithelial cells Dong Jin-hui, Ren Xiu-min, Xu Ou, Wang Jian-xing 2016, 20 (32):  4764-4770.  doi: 10.3969/j.issn.2095-4344.2016.32.008 Abstract ( 372 )   PDF (4231KB) ( 300 )   Save BACKGROUND: Damage to nasal ciliated epithelial cells can lead to a severe injury in nasal biological function. Compared with other adult stem cells, human umbilical cord blood stem cells have better differentiation potential. OBJECTIVE: To explore the feasibility of human umbilical cord blood stem cells differentiating into nasal ciliated epithelial cells through in vitro culture and induction techniques. METHODS: Normal and healthy umbilical cord blood samples were collected to isolate human umbilical cord blood stem cells, followed by identification and subculture in vitro. Umbilical cord blood stem cells at passage 3 were infected with recombinant adeno-associated virus carrying enhanced green fluorescent protein and cultured using air liquid interface culture method. Thereafter, PCR assay was employed for detecting MUCS expression in cultured stem cells at 1 and 2 weeks after induction, and immunofluorescent staining for FOXJ1 was performed at 3 weeks. RESULTS AND CONCLUSION: After subculture, passage 3 umbilical cord blood stem cells that could express stem cell surface markers were visible in a uniform shape and had good refraction. After 3 hours of gene transfection, green fluorescence issued from the passage 3 cells were visible, and the cell positive rate was up to 96.2% until 48 hours, indicating good transfection efficiency. RT-PCR findings showed that MUC8 mRNA had no expression in the umbilical cord blood stem cells, but expressed strongly in the nasal ciliated epithelial cells, whose expression was weak at 1 week of culture and increased at 2 weeks. Additionally, the positive expression of FOXJ1 red fluorescence was observed under the transfection of green fluorescent protein. These results suggest that human umbilical cord blood stem cells could differentiate into nasal epithelial cells under suitable conditions.     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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Leonurine combined with bone marrow mesenchymal stem cell transplantation in the treatment of intrauterine adhesions Wang Bing-yu, Ji Xia 2016, 20 (32):  4771-4777.  doi: 10.3969/j.issn.2095-4344.2016.32.009 Abstract ( 302 )   PDF (5902KB) ( 229 )   Save BACKGROUND: Increasing evidence has demonstrated that bone marrow mesenchymal stem cell (BMSC) transplantation can promote skin, liver and lung repair in animal models; and Leonurus sibiricus L. has the ability of promoting blood circulation and regulating menstruation. OBJECTIVE: To investigate the therapeutic effect of BMSC transplantation with leonurine on intrauterine adhesions (IUA) in rabbits and the relevant mechanism of action. METHODS: After modeled using dual injury method, rabbit models of IUA were randomly divided into five groups: sham-operated group (sham), model group (IUA), BMSC transplantation group, leonurine treatment group and combined treatment group (BMSCs+leonurine). Rabbits in the sham group were only given normal saline rinsing after hysterotomy, while those in the latter three groups were correspondingly given intrauterine BMSC transplantation or/and intragastric administration of 4 mg/kg leonurine for 14 days. Morphological changes of the endometrium were observed using hematoxylin-eosin staining, and expression levels of transforming growth factor β protein, Smad3 protein and interferon-γ mRNA were detected using immunohistochemical staining and real-time fluorescence quantitative PCR, respectively. RESULTS AND CONCLUSION: Compared with the model group, the degree of IUA was all significantly improved in the other groups, especially in the combined treatment group. Moreover, BMSC transplantation, leonurine treatment and their combined use all could inhibit IUA-induced increase of transforming growth factor β and Smad3 protein expression and IUA-induced decrease of interferon-γ mRNA level. Importantly, all these alternations were much more pronounced in the combined treatment group. Our results show that the combined use of BMSC transplantation and leonurine treatment can exert a synergistic effect in the improvement of IUA through the transforming growth factor β/Smad3 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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Effect of lovastatin on proliferation and apoptosis of glioma stem cells Wen Gong-ling, Wen Chang-ming, Wang Yan-ping, Kang Mei-juan, Zhou Jing, Zhang Bao-chao 2016, 20 (32):  4778-4784.  doi: 10.3969/j.issn.2095-4344.2016.32.010 Abstract ( 213 )   PDF (1074KB) ( 259 )   Save BACKGROUND: Increasing evidence has shown that lovastatin with less toxicity to normal cells has crucial effects on proliferation, apoptosis and differentiation of various cancer cells. However, its roles in glioma stem cells remain unclear. OBJECTIVE: To explore the effect of lovastatin on proliferation and apoptosis of glioma stem cells. METHODS: Flow cytometric sorting was used to separate glioma stem cells from human glioblastoma cell line U87. Effects of lovastatin on the proliferation and apoptosis of glioma stem cells were determined by MTT and flow cytometry, respectively. Furthermore, expression levels of Ki67, Bax and Bcl-2 in glioma stem cells treated with lovastatin were detected using western blot analysis. RESULTS AND CONCLUSION: The CD133-positive glioma stem cells were sorted from human glioblastoma cell line U87 with a positive percentage of 85%. MTT assay showed that lovastatin inhibited the proliferation of glioma stem cells in dose (5, 10, 20 μmol/L)- and time (24, 48, 72, 96 hours)-dependent manners. Flow cytometry analysis showed that 10 μmol/L lovastatin (48 hours) induced apoptosis in glioma stem cells. In addition, the expression level of Ki67 was decreased by lovastatin treatment in a dose-dependent manner, and the Bcl-2 and Bax expression levels were reduced and increased by 10 μmol/L lovastatin treatment, respectively. In conclusion, lovastatin can inhibit cell proliferation and induce apoptosis of glioma stem cells, and lovastatin may be a potential drug for treatment of brain tumors.     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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Epimedium combined with bone marrow mesenchymal stem cell transplantation for treatment of kidney disease in rats Chen Xiao-yong, Wang Zi-chuang, Zhang Jian-wei, Han Jia-rui 2016, 20 (32):  4785-4791.  doi: 10.3969/j.issn.2095-4344.2016.32.011 Abstract ( 255 )   PDF (5420KB) ( 207 )   Save BACKGROUND: A large number of studies have shown that bone marrow mesenchymal stem cells (BMSCs) with differentiation ability can be used to treat kidney injury. Epimedium has good anti-inflammatory and immune restoration ability. OBJECTIVE: To observe the effect of epimedium with BMSC transplantation on rat’s kidney disease caused by adriamycin nephrosis. METHODS: Fifty rats were equivalently randomized to receive no treatment in control group, tail vein injection of normal saline plus intragastric injection of normal saline in model group, tail vein injection of BMSCs plus intragastric injection of epimedium decoction in combined group, tail vein injection of BMSCs in BMSCs group, and intragastric injection of epimedium decoction in epimedium group. Rat models of adriamycin nephrosis were made in the latter four groups through tail vein injection of doxorubicin hydrochloride, while rats in the control group were only given the same volume of normal saline. Twenty-four hours after modeling, rats in the latter four groups began to be given the corresponding treatments. Epimedium decoction was given in the combined and epimedium groups at a dose of 1 mL/d, for consecutive 1 week.  RESULTS AND CONCLUSION: Compared with the model group, mitigated kidney injury and reduced urinary protein level were found in rats undergoing BMSCs and/or epimedium decoction treatment; the levels of plasma albumin and catalase were significantly increased in the combined group and epimedium group, while the levels of serum cholesterol, triglycerides and malondialdehyde decreased; the mRNA expression of transforming growth factor β1 in the kidney tissues was dramatically declined in the combined group and BMSC group. To conclude, the combined use of epimedium and BMSCs is superior to their use alone in the treatment of kidney disease.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cells for repairing acute respiratory distress syndrome Zhang Lei 2016, 20 (32):  4792-4797.  doi: 10.3969/j.issn.2095-4344.2016.32.012 Abstract ( 411 )   PDF (4175KB) ( 187 )   Save BACKGROUND: To date little is reported clinically on bone marrow mesenchymal stem cells (BMSCs) for acute respiratory distress syndrome (ARDS). OBJECTIVE: To study the effect of BMSCs in ARDS rats. METHODS: Sixty-three Sprague-Dawley rats were randomly assigned into normal control group, model and BMSCs groups (n=21 per group), followed by establishment of ARDS models. After awakening, rats in the model group were given no treatment, while those in the BMSCs group given tail vein injection of fluorouracil-labeled BMSCs. Then, pathological observation of the lung tissue was conducted using hematoxylin-eosin staining, and ELISA method was employed to detect interleukin-1β level in the rat lung tissue homogenates. Repair effects on lung injury were compared between two groups. RESULTS AND CONCLUSION: Hematoxylin-eosin staining showed that inflammation, edema and congestion in the rat lung tissue were vanished gradually, but no consolidation was found in the BMSCs group, while there was visible edema and congestion in the lung tissue of rats in the model group. Compared with the model group, pulmonary edema and interleukin-1β level in the lung tissue were significantly reduced in the BMSCs group (P < 0.05). Therefore, the whole bone marrow culture method is suitable to obtain BMSCs that have desired effects on ARDS in rats. In ARDS rats, due to inhalation of lipopolysaccharide, BMSCs can be directly involved in lung epithelial repair by reducing the release of inflammatory factors.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of bone marrow mesenchymal stem cell transplantation on senile dementia behaviors Gao Ming-long, Sun Li, Zhao Xiao-chuan, Yu Ming, Wang Jin-cheng 2016, 20 (32):  4798-4804.  doi: 10.3969/j.issn.2095-4344.2016.32.013 Abstract ( 257 )   PDF (5638KB) ( 207 )   Save BACKGROUND: Drug treatment for senile dementia has unsatisfactory outcomes although to a certain extent it can reduce and delay the progression of Alzheimer’s disease. Stem cell transplantation is a new attempt for the treatment of senile dementia. OBJECTIVE: To observe the effect of bone marrow mesenchymal stem cell transplantation on the behavior of senile dementia rats. METHODS: Rat models of senile dementia were made in 20 Sprague-Dawley rats that were given continuous 60-day gavage of aluminium chloride solution. Then, model rats were randomized into model group treated with normal saline injection and experimental group treated with hippocampal injection of bone marrow mesenchymal stem cells, respectively. Another 10 rats undergoing normal feeding served as control group. Learning and memory ability of rats were tested by Morris water maze, and superoxide dismutase activity and malondialdehyde content in brain tissues of rats were measured by colorimetric method at 4 weeks after cell transplantation. RESULTS AND CONCLUSION: Compared with the model group, the escape latency was shortened and the cross-platform frequency was increased in the experimental group (P < 0.05), and moreover, significantly elevated superoxide dismutase activity and reduced malondialdehyde content in the brain tissues of rats were found in the experimental group (P < 0.05). These findings indicate that bone marrow mesenchymal stem cell transplantation contributes to behavior improvement in senile dementia rats by improving the learning and memory ability.     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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Embryonic neural stem cell transplantation for Alzheimer’s disease Zhao Hui 2016, 20 (32):  4805-4810.  doi: 10.3969/j.issn.2095-4344.2016.32.014 Abstract ( 288 )   PDF (3745KB) ( 497 )   Save BACKGROUND: More recently, stem cell therapy has become an issue of concern. Exogenous neural stem cell transplantation brings new hope for the treatment of nervous system injury by self-replication and differentiation to complement and replace damaged or dead nerve cells. OBJECTIVE: To explore the therapeutic efficacy of neural stem cell transplantation on Alzheimer’s disease. METHODS: Thirty APP/PS1 mice with Alzheimer’s disease were randomly assigned into model group, cell solution transplantation group or cell transplantation group (n=10 per group). Another 10 C57BL/6 mice were selected as controls. Embryos of C57BL/6 mice at 18 embryonic days were taken to make neural stem cell suspension followed by transfection using lentiviral vectors carrying GFP gene at different multiplicities of infection (1, 5, 10, 15, 20). Afterwards, GFP-transfected neural stem cells were implanted into the hippocampus of Alzheimer’s disease mice in the cell transplantation group, while the same volume of complete medium was injected into the hippocampus of mice in the cell solution transplantation group. Morris water maze test was performed at 2 weeks after cell transplantation, and brain tissues of mice was taken and detected histologically at 4 weeks after cell transplantation. RESULTS AND CONCLUSION: Compared with the control group, the escape latency was significantly higher, and the number of crossings over the target quadrant was lower in the other three groups (P < 0.05). Compared with the cell solution transplantation and model groups, in contrast, the escape latency was significantly lower, and the number of crossings over the target quadrant was significantly higher in the cell transplantation group (P < 0.05). Four weeks after transplantation, more intact neurons were found in the cell transplantation group as compared with the model group. These findings indicate that neural stem cell transplantation can improve behavior and morphology performance of mice with Alzheimer’s disease.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Adipose-derived mesenchymal stem cell transplantation for treatment of liver failure Hu Xiao-yang, Leng Hui-guo, Mao Cui-yan, Zhang Yuan 2016, 20 (32):  4811-4817.  doi: 10.3969/j.issn.2095-4344.2016.32.015 Abstract ( 255 )   PDF (1110KB) ( 250 )   Save BACKGROUND: Studies have found that both bone marrow mesenchymal stem cells and umbilical cord mesenchymal stem cells have therapeutic effects on liver failure, but little is reported on adipose-derived mesenchymal stem cell transplantation for treatment of liver failure. OBJECTIVE: To discuss the therapeutic effect of adipose-derived mesenchymal stem cell transplantation on rat liver failure. METHODS: Forty-five Sprague-Dawley rats were equally randomized into transplantation, model and control group. Except the control group, animal models of liver failure were made by intraperitoneal injection of D-galactosaminein in the transplantation and model groups. One week after modeling, rats in the transplantation group were given 1 mL of adipose-derived mesenchymal stem cells via the tail vein. RESULTS AND CONCLUSION: Adipose-derived mesenchymal stem cell transplantation successfully upregulated alanine aminotransferase and aspartate aminotransferase levels, and moreover, neatly arranged liver cells, reduced inflammatory cells and intact hepatic lobule were shown on B ultrasound after cell transplantation. All these findings indicate that adipose-derived mesenchymal stem cell transplantation has therapeutic effects on rat liver failure by improving rat’s liver function and pathological changes.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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High-dose cyclophosphamide-induced immunogenic tolerance following haploidentical allogeneic hematopoietic stem cell transplantation in severe aplastic anemia children Guo Zhi, Tong Chun, Liu Xiao-dong, Yang Kai, He Xue-peng, Zhang Yuan, Chen Peng, Lou Jin-xing, Chen Hui-ren 2016, 20 (32):  4818-4824.  doi: 10.3969/j.issn.2095-4344.2016.32.016 Abstract ( 371 )   PDF (1213KB) ( 242 )   Save BACKGROUND: High-dose cyclophosphamide (CTX)-induced immunogenic tolerance following haploidentical allogeneic hematopoietic stem cell transplantation (allo-HSCT) is developed to optimize the treatment of childhood severe aplastic anemia (SAA) using haplotype allo-HSCT, providing a theoretical basis for the clinical application. OBJECTIVE: To investigate the clinical efficacy and safety of the use of high-dose CTX following haploidentical allo-HSCT in SAA children. METHODS: Clinical data from 10 children with SAA undergoing haploidentical allo-HSCT at the Department of Hematology, General Hospital of Beijing Military Area from January 2013 to January 2015 were retrospectively analyzed. Pretreatment was CTX, fludarabine, Busulfex combined with anti-human lymphocyte immune globulin used for 2 consecutive days, and then 3 days after transplantation, CTX (50 mg/kg per day) was used to induce immunogenic tolerance. Combined use of cyclosporin A, methotrexate and tacrolimus functioned as a prophylaxis for graft-versus-host disease. Another 10 SAA children who underwent synchronous HLA-identical sibling HSCT served as controls. Complications and survival in children were statistically analyzed in the two groups. RESULTS AND CONCLUSION: In the treatment group, children were followed up until May 2015, and the median follow-up period was 18.1 months (5-28 months). Hematopoietic reconstruction was successful in all cases, and there were three cases of graft-versus-host disease, three cases of pulmonary infection and two cases dying of pulmonary infection. In the control group, the median follow-up period was 20.7 months (6-27 months), and all the children received hematopoietic reconstruction. Additionally, there were two cases of graft-versus-host disease, four cases of pulmonary infection, one case dying of graft-versus-host disease and one case dying of pulmonary infection in the control group. The total survival rate in each group was 80%. In summary, high-dose CTX-induced immunogenic tolerance is safe and effective for SAA children undergoing haploidentical allo-HSCT, which makes the clinical efficacy of haploidentical allo-HSCT identical to that of matched HSCT.   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 stem cell transplantation for osteoporosis rats: evaluation by bone mineral density and histomorphometry measurement Li Guang-zhang, Jiang Zhao-qin, Liu Zhi-xin 2016, 20 (32):  4825-4830.  doi: 10.3969/j.issn.2095-4344.2016.32.017 Abstract ( 278 )   PDF (4274KB) ( 204 )   Save BACKGROUND: Stem cell transplantation is increasingly hoped to promote osteoblast differentiation and inhibit osteoclast proliferation in the treatment of osteoporosis. OBJECTIVE: To study the therapeutic effect of exogenous adipose-derived stem cell (ADSC) transplantation on osteoporosis in ovariectomized rats. METHODS: Thirty Sprague-Dawley female rats were equivalently randomized into sham, model, ADSC transplantation groups. Rats in all groups except the sham group underwent bilateral ovariectomy to make osteoporosis models. Surrounding adipose tissues instead of the ovary were removed in the sham group. After modeling, rats were given 2×106 ADSCs at passage 4 via the tail vein in the transplantation group and the same volume of normal saline in the model group, once a week. After 6 weeks, levels of serum calcium, phosphorus, and alkaline phosphatase as well as bone mineral density and histomorphometry indicators were detected in rats. RESULTS AND CONCLUSION: Compared with the sham group, the trabecular bone volume fraction was significantly decreased in the model group (P < 0.01), but remarkably increased after ADSC transplantation (P < 0.05). After modeling, the bone trabecular absorption surface percentage and rate of bone trabecular formation were elevated significantly (P < 0.05 or P < 0.01), while these increases were improved by ADSC transplantation (P < 0.05). Additionally, the levels of serum calcium and alkaline phosphatase and bone mineral density were significantly decreased after modeling, but were increased after ADSC transplantation. In contrast, the serum level of phosphorus was significantly increased in the model group (P < 0.05) but decreased markedly in the ADSC transplantation group (P < 0.05). To conclude, ADSC transplantation can reduce the loss of bone mass in osteoporosis rats by ovariectomy.   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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Tumor necrosis factor-alpha pretreated umbilical cord blood mesenchymal stem cell transplantation for treatment of myocardial infarction Wang Wei, Li Xiao-fu, Li Zhong-jian 2016, 20 (32):  4831-4837.  doi: 10.3969/j.issn.2095-4344.2016.32.018 Abstract ( 279 )   PDF (4706KB) ( 215 )   Save BACKGROUND: A large amount of inflammatory mediators from myocardial cells are secreted in  response to myocardial injury after myocardial ischemia. Moreover, inflammatory cytokines in infarction and ischemia regions contribute to myocardial tissue repair and adaptation. OBJECTIVE: To investigate the effect of tumor necrosis factor-α (TNF-α) pretreatment on cardiac function of myocardial infarction rabbits undergoing umbilical cord blood mesenchymal stem cell transplantation. METHODS: Thirty-six white rabbits were equally randomized into sham, model, non-TNF-α, and TNF-α groups. Animal models of myocardial infarction were made in the latter three groups. Twenty-four hours after modeling, PBS, umbilical cord blood mesenchymal stem cells pretreated with or without TNF-α were injected at infarct center and border, respectively, in the model, TNF-α and non-TNF-α groups. RESULTS AND CONCLUSION: Model rabbits in the TNF-α and non-TNF-α groups showed better cardiac function and lower size of myocardial infarction and fibrosis than those in the model group. Compared with the non-TNF-α group, moreover, the TNF-α group showed better outcomes in these indicators. These findings indicate that TNF-α pretreatment can markedly improve the therapeutic efficacy of umbilical cord blood mesenchymal stem cell transplantation on myocardial infarction.   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 cell transplantation for the treatment of chronic kidney disease in rats Zhao Ya-pei, Liu Cui-jing, Yang Cui-ying, Fan Shi-ying, Zhang Li-jian, Zhao Yan-qin, Xie Ying-xin 2016, 20 (32):  4838-4844.  doi: 10.3969/j.issn.2095-4344.2016.32.019 Abstract ( 301 )   PDF (4014KB) ( 229 )   Save BACKGROUND: Urine-derived stem cells are most likely to come from the kidney tissue, and therefore, these cells are more adaptable to kidney microenvironment, providing a new option for the treatment of kidney diseases. OBJECTIVE: To explore the therapeutic efficacy of human urine-derived stem cells on chronic nephropathy rats. METHODS: The fresh urine samples of healthy people were collected, and then human urine-derived stem cells were extracted and cultured in vitro. Twenty Sprague-Dawley rats were used to prepare chronic nephropathy models, and given injection of human urine-derived stem cell suspension (experimental) or normal saline (control) into the renal cortex, respectively. Another 10 healthy rats were used as controls. Therapeutic effects on renal function were assessed by detection of serum creatinine level and glomerular filtration rate in the three groups. The kidney tissues of rats were taken and observed histomorphologically in each group. RESULTS AND CONCLUSION: Human urine-derived stem cells were found to remarkable improve rat’s renal function as well as reduce the histomorphological changes in the kidney tissues of rats. Compared with the control group, the serum creatinine level was decreased while the glomerular filtration rate was increased significantly in the experimental group; CD68 expression and infiltration of interstitial inflammatory cells were also markedly reduced in the experimental group. To conclude, human urine-derived stem cells can improve the renal function of chronic nephropathy 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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Effects of drug-contained sera of Naoluo Xintong versus Zuogui pill on the proliferation and differentiation of in vitro cultured neural stem cells Chen Li-ming, Pei Lei 2016, 20 (32):  4845-4851.  doi: 10.3969/j.issn.2095-4344.2016.32.020 Abstract ( 395 )   PDF (5145KB) ( 192 )   Save BACKGROUND: Because of limited source and a relatively weak ability of differentiation and proliferation, how to take positive and effective measures to promote neural stem cell proliferation, differentiation has become the focus of research. OBJECTIVE: To investigate the effects of drug-contained sera of Naoluo Xintong versus Zuogui pill on the proliferation and differentiation of in vitro cultured rat neural stem cells. METHODS: Embryonic neural stem cells of Sprague-Dawley rats were isolated and cultured in vitro, and then were co-cultured with the serum medium containing 10% Naoluo Xintong and 10% Zuogui pill, respectively. Comparative observations were performed between two groups by inverted microscope and immunofluorescence staining. RESULTS AND CONCLUSION: Under the inverted microscope, the cells began to grow in cluster and gather into a ball, but the diameter was relatively small after 24-hour culture; the neurospheres expended further, with relatively regular shape, but no neurosphere differentiation appeared after 48 hours of culture. The average prominent length of the neurospheres in the Zuogui pill group was significantly greater than that in the Naoluo Xintong group after 5 days of culture (P < 0.05). The rate of rat neural stem cells differentiating into MAP-2-positive cells in the Zuogui pill group was significantly lower than that in the Naoluo Xintong group (P < 0.05), but the rate of differentiated cells positive for glial fibrillary acidic protein in the Zuogui pill group was significantly higher than that in the Naoluo Xintong group (P < 0.05). After 48 hours of culture, neurospheres were cultured in the different drug-contained media, and 12 hours later, the neurospheres adhered to the wall, and a small amount of cell migration occurred. Then, cell migration began to increase with time. Under the immunofluorescence staining: prominent neurons with long protrusions were increased in both two groups, but there were no significant differences in the proportion of neurons and astrocytes between two groups (P > 0.05). These findings suggest that drug-contained sera of Naoluo Xintong and Zuogui pill can both not only promote the proliferation and differentiation of in vitro cultured rat neural stem cells, but also provide a suitable microenvironment for neural cell proliferation. Additionally, there are significant differences between the two drugs. Consequently, it is feasible to induce neural cell proliferation and differentiation by Naoluo Xintong and Zuogui pill.   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 mesenchymal stem cells from psoriatic skin versus normal skin Zhang Jing, Wang Sheng, Kang Kang, Wang Jian-mei 2016, 20 (32):  4852-4858.  doi: 10.3969/j.issn.2095-4344.2016.32.021 Abstract ( 322 )   PDF (4375KB) ( 210 )   Save BACKGROUND: Skin-derived mesenchymal stem cells may reflect the onset of psoriasis. OBJECTIVE: To analyze the biological characteristics of skin-derived mesenchymal stem cells in psoriasis patients. METHODS: Skin-derived mesenchymal stem cells from 30 patients with psoriasis and 20 healthy controls were isolated and cultured by trypsin. Flow cytometry was used to detect the cellular immune phenotypes CD34, CD44, CD29, CD45, CD90, CD105, CD73 and HLA-DR. The mesenchymal stem cells were induced by the corresponding cartilage, osteogenic and osteogenic inducing agents, to identify the multi-directional differentiation ability. The cell proliferation curve was plotted at passage 3, and the levels of transforming growth factor-β1 and epidermal growth factor in culture supernatant were detected by ELISA assay. RESULTS AND CONCLUSION: Under an inverted phase contrast microscope, primary skin-derived mesenchymal stem cells isolated from patients with psoriasis and normal controls both exhibited heterogeneity. In the two groups, CD29, CD90, CD44, CD73 and CD105 were highly expressed, and CD45, CD34, and HLA-DR were lowly expressed. Under certain conditions, skin-derived mesenchymal stem cells were induced to differentiate into adipocytes, osteoblasts or chondrocytes. Proliferation of skin-derived mesenchymal stem cells in the psoriasis group was significantly faster than that in control group, but the final number of cells in the two groups tended to be consistent. The levels of transforming growth factor-β1 and epidermal growth factor in the psoriatic skin had no correlation with the severity of the disease (P > 0.05). Compared with the control group, the epidermal growth factor level in the cell supernatant was significantly higher in the psoriasis group (P < 0.01), while the level of transforming growth factor-β1 was significantly lower (P < 0.01). These results showed that there is heterogeneity in the morphology of skin-derived mesenchymal stem cells from psoriasis patients, and the biological activity of mesenchymal stem cells is abnormal.   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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Electroacupuncture promotes the proliferation and differentiation of neural stem cells in the rat hippocampus of spleen deficiency syndrome Miao Ling 2016, 20 (32):  4859-4864.  doi: 10.3969/j.issn.2095-4344.2016.32.022 Abstract ( 358 )   PDF (1027KB) ( 168 )   Save BACKGROUND: Spleen deficiency has varying impact on the immune system and digestive system of the human body, and can also damage the normal function of the central nervous system as there is a close relationship between the spleen and brain. OBJECTIVE: To explore the effect of electroacupuncture on the proliferation and differentiation of neural stem cells in the rat hippocampus of spleen deficiency syndrome. METHODS: Sixty Sprague-Dawley rats were randomly divided into three groups: normal, control and electroacupuncture groups. The animal model of spleen deficiency syndrome was prepared in the control group and electroacupuncture group. Two weeks after modeling, rats in the electroacupuncture group were given electroacupuncture treatment, and the changes of body mass in normal rats and model rats were measured. Five rats from each group were taken to observe the histological changes of neural stem cells in the hippocampus at 1, 2, 3, 4 weeks after treatment. RESULTS AND CONCLUSION: The body mass of rats in the model group was significantly lower than that in the normal group at 2 weeks after modeling (P < 0.05). In the electroacupuncture group, the number of BrdU positive cells was significantly higher than that in the normal group and control group at 2, 3, 4 weeks after modeling (P < 0.05); the number of BrdU/Nestin positive cells was significantly higher than that in the normal group at 2, 3, 4 weeks (P < 0.05); the number of BrdU/glial fibrillary acidic protein positive cells was significantly higher than that in the normal group at 2 weeks (P < 0.05); and the number of BrdU/neuronspecific enolase positive cells was significantly higher than the normal group at 3 weeks (P < 0.05). In the control group, the number of BrdU/Nestin positive cells was significantly lower than that in the normal group and electroacupuncture group at 1 week after modeling, significantly lower than that in the electroacupuncture group at 2 and 4 weeks, and significantly lower than the normal group at 3 weeks (P < 0.05); the number of BrdU/glial fibrillary acidic protein positive cells was significantly lower than that in the normal group and electroacupuncture group at 1, 2, 4 weeks; and the number of BrdU/neuronspecific enolase positive cells was significantly lower than that in the normal group and electroacupuncture group at 1, 2 weeks. Taken together, these experimental findings show that electroacupuncture treatment effectively promotes the proliferation of hippocampal neural stem cells that are induced to differentiate into astrocytes and neurons.   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