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    01 April 2016, Volume 20 Issue 14 Previous Issue    Next Issue
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    Combined use of interleukin-6 receptor monoclonal antibody and bone marrow mesenchymal stem cells reduces neuronal apoptosis after acute spine cord injury
    Ren Yi-xing, Meng Xian-yong, Hu Chang-bo, Yang Xin-ming
    2016, 20 (14):  1981-1988.  doi: 10.3969/j.issn.2095-4344.2016.14.001
    Abstract ( 209 )   PDF (1358KB) ( 542 )   Save

     BACKGROUND: Studies have suggested that interleukin-6 is crucial for inducing cell apoptosis after acute spinal cord injury.
    OBJECTIVE: To observe the effect of interleukin-6 receptor monoclonal antibody combined with bone marrow mesenchymal stem cells to treat acute spinal cord injury in rats.
    METHODS: Thirty Sprague-Dawley rats were randomly divided into sham group, model group (spinal cord injury group), treatment group 1 (interleukin-6 receptor monoclonal antibody transplantation group), treatment group 2 (bone marrow mesenchymal stem cell transplantation group), treatment group 3 (bone marrow mesenchymal stem cell+interleukin-6 receptor monoclonal antibody group), with six rats in each group. In the sham group, the spinal cord was only exposed with no injury, and in the other four groups, rat models of acute spinal cord injury were made using modified Allen’s method. Local injection treatment was performed in all the groups at 28 days after modeling. Basso, Beattie and Bresnahan (BBB) scoring and improved Tarlov scoring were used at 1 day before treatment and 1, 3, 7, 14, 28 days after treatment to test the hindlimb function. At 28 days after treatment, TUNEL method was used to detect cell apoptosis in the spinal cord.
    RESULTS AND CONCLUSION: Compared with the sham group, BBB scores and improved Tarlov scores were decreased significantly in the other four groups (P < 0.05). At 7 days after treatment, the BBB scores and improved Tarlov scores in the treatment group 3 were significantly higher than those in the model group (P < 0.05). At 14 days after treatment, the BBB scores and improved Tarlov scores in the treatment groups 1 and 2 were significantly higher than those in the model group (P < 0.05); compared with the treatment group 2, the BBB score and improved Tarlov score were significantly increased in the treatment group 3 (P < 0.05). Compared with the sham group, the number of apoptotic cells was significantly increased in the other four groups (P < 0.05); compared with the model group, the number of apoptotic cells was significantly decreased in the three treatment groups (P < 0.05); compared with the treatment group 2, the number of apoptotic cells was significantly lower in the treatment group 3 (P < 0.05). These findings indicate that the combined use of interleukin-6 receptor monoclonal antibody and bone marrow mesenchymal stem cell transplantation is better than bone marrow mesenchymal stem cell transplantation alone in the treatment of spinal cord injury, and interleukin-6 receptor monoclonal antibody reduces cell apoptosis in spinal cord injury, which is of positive significance for preventing against acute spinal cord injury.

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    Hypoxic preconditioning inhibits apoptosis of bone marrow mesenchymal stem cells through overexpressing Pim-1
    Zhang You, Yan Wei-ya, Shen Zheng-ya, Yang Jun-jie, Hui Jie
    2016, 20 (14):  1989-1998.  doi: 10.3969/j.issn.2095-4344.2016.14.002
    Abstract ( 260 )   PDF (6710KB) ( 760 )   Save

    BACKGROUND: Bone marrow mesenchymal stem cells have a low survival rate after implanted into the ischemic myocardium. However, hypoxia preconditioning (HPC) may enhance bone marrow mesenchymal stem cell proliferation and promote its survival rate.
    OBJECTIVE: To explore whether Pim-1 is involved in HPC protecting against apoptosis of bone marrow mesenchymal stem cells and the relevant mechanism.
    METHODS: Bone marrow mesenchymal stem cells were respectively subjected to HPC for 0, 6, 12, and 24 hours. The expression of Pim-1 and apoptosis-related genes were detected by RT-qPCR and western blot. Then, the best hypoxic preconditioning time was determined as 12 hours. Then, bone marrow mesenchymal stem cells were assigned to one of the following groups: control (without HPC), 12-hour HPC, 12-hour HPC+Pim-1 inhibitor groups. Flow cytometry analysis was used to detect the cell apoptosis, Transwell assay to analyze the cell migration ability in each group, and JC-1 kit to detect mitochondrial membrane potential. Animal models of myocardial infarction were established. One week after modeling, bone marrow mesenchymal stem cells were given via multi-point injection around the infarct zone of rats. Two weeks after modeling, heart tissues of rats were taken and sliced followed by DiI staining to calculate the survival rate of bone marrow mesenchymal stem cells. Additionally, rat cardiac function was assessed by echocardiography prior to and after modeling as well as at 4 weeks after cell transplantation.
    RESULTS AND CONCLUSION: At 12 hours after HPC, the expression of Pim-1, p-Akt and Bcl-2 gene in the infarct region was significantly increased, but the expression of caspase-3 and Bax was significantly decreased. Compared with the control group, cell viability in the 12-hour HPC group was increased very significantly at 1 week after cell transplantation (P < 0.001), the migration and anti-apoptosis ability were enhanced significantly (P < 0.01) and the cardiac function of rats was significantly improved in the 12-hour HPC group (P < 0.05). All of these protective effects were blocked by the Pim-1 inhibitor. These findings indicate that HPC can protect bone marrow mesenchymal stem cells from apoptosis through activating Akt and up-regulating Pim-1, and thereby improve the therapeutic effect of bone marrow mesenchymal stem cell transplantation on ischemic heart diseases.
    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 

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    Indirect co-culture with endothelial progenitor cells improves proliferation and apoptosis of bone marrow mesenchymal stem cells of osteoporosis rats
    Liu Zhu-ying, Chen Ying, Liu Qian, Liang Yuan, Zhang Rui, Wen Yi, Ding Yin
    2016, 20 (14):  1999-2006.  doi: 10.3969/j.issn.2095-4344.2016.14.003
    Abstract ( 305 )   PDF (7289KB) ( 569 )   Save

    BACKGROUND: Previous studies have found that estrogen deficiency causes a reduction in the activity of bone marrow mesenchymal stem cells (BMSCs).
    OBJECTIVE: To explore the effect of endothelial progenitor cells (EPCs) on the BMSCs proliferation and apoptosis ability of osteoporosis rats.
    METHODS: Healthy female Sprague-Dawley rats, 6 weeks old, were enrolled and subjected to bilateral ovariectomy to make osteoporosis models. BMSCs and EPCs were isolated using density gradient centrifugation combined with adhesion method, and identified with surface markers, cell proliferation and immunocytochemistry in vitro. We used Transwell inserts to establish EPCs and OVX-BMSCs indirect co-culture system. Control groups were OVX-BMSCs group and sham-BMSCs group in which rats were only subjected to remove the equal amount of fat tissues around the ovary. Flow cytometry was applied to detect BMSCs proliferation and apoptosis ability.
    RESULTS AND CONCLUSION: Compared with the control groups, the results of flow cytometry test showed that the proportion of OVX-BMSCs at S phase was significantly increased at 3 days of indirect co-culture with EPCs and the apoptosis rate was significanty reduced at 10 days of indirect co-culture with EPCs (both P < 0.05). These results suggest that EPCs can promote the proliferation but inhibit the apoptosis of OVX- BMSCs.
    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 

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    Iron overload inhibits osteogenesis and promotes adipogenesis in human bone marrow mesenchymal stem cells by producing reactive oxygen
    Han Yan-jiu, Liu Guo-hui, Liu Yong
    2016, 20 (14):  2007-2014.  doi: 10.3969/j.issn.2095-4344.2016.14.004
    Abstract ( 369 )   PDF (4802KB) ( 390 )   Save

    BACKGROUND: Iron overload as a new etiological factor participate in the pathogenesis of senile osteoporosis, but its mechanism remains unclear. But the mechanism of iron overload cause senile osteoporosis pathogenesis remains unclear. Osteogenic/adipogenic differentiation homeostasis of bone marrow mesenchymal stem cells (BMSCs) plays an important role in the maintenance of normal human bone mass, and its imbalance can lead to senile osteoporosis. Iron effects on the osteogenic/adipogenic differentiation of BMSCs remain unclear. Therefore, we clarify the role of iron overload in the pathogenesis of senile osteoporosis by studying the effect of iron overload on BMSCs osteogenic/adipogenic differentiation.
    OBJECTIVE: To explore the effect of iron overload on the osteogenesis and adipogenesis of human BMSCs (hBMSCs) in vitro and to explicit the role of iron overload in the pathogenesis of senile osteoporosis.
    METHODS: hBMSCs were isolated by using density gradient centrifugation method. Isolated cells were treated with normal saline as normal control group, treated with 10, 20, 30 μmol/L ferrous citrate to mimic iron overload conditions, or treated with 30 μmol/L ferrous citrate followed by 0.5 mmol/L N-acetylcysteine as an antioxidant.
    RESULTS AND CONCLUSION: Iron overload did not remarkably inhibit the proliferation of hBMSCs, but it could promote the generation of reactive oxygen in hBMSCs. After treatment with ferrous citrate, the expression of c-Maf and Runx2 decreased, and the expression of peroxisome proliferator-activated receptor γ increased; moreover, formation of calcium nodules decreased, but lipid droplets were produced. N-acetylcysteine could inhibit the production of reactive oxygen and increase the expression of c-Maf induced by iron overload. These findings indicate that iron overload inhibits osteogenesis and promotes adipogenesis in hBMSCs through the generation of reactive oxygen. Meanwhile, c-Maf plays an essential role in the iron-overload induced differetiation imbalance.
    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 

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    Bone marrow mesenchymal stem cell transplantation for repair of mesangial proliferative glomerulonephritis: role and mechanism
    Liao Dan, Zhang Lin, Dai Xiao-yu, Wang Jia-li, Du Xiao-jiong
    2016, 20 (14):  2015-2020.  doi: 10.3969/j.issn.2095-4344.2016.14.005
    Abstract ( 319 )   PDF (3853KB) ( 373 )   Save

    BACKGROUND: Bone marrow mesenchymal stem cells are likely to repair renal injury by differentiating into renal parenchymal cells.
    OBJECTIVE: To explore the effect and mechanism of bone marrow mesenchymal stem cells in the renal repair after mesangial proliferative glomerulonephritis.
    METHODS: Thirty Sprague-Dawley rats were randomized into normal group, model group and treatment group (n=10 per group). Model group and treatment group were treated with tail vein injection of mouse anti-rat monoclonal antibody Thy1.1 to prepare mesangial proliferative glomerulonephritis models. One week after modeling, rats in the treatment group were given 2×106 bone marrow mesenchymal stem cells via the tail vein, and rats in the other two groups were given the same volume of normal saline. Two weeks after transplantation, urinary protein, urea nitrogen, creatinine levels were detected; hematoxylin-eosin staining was used for observing pathological changes of the renal tissue under microscope; and the expression of transforming growth factor beta 1 was detected by immunohistochemistry method.
    RESULTS AND CONCLUSION: The levels of urinary protein, urea nitrogen and creatinine as well as the expression of transforming growth factor beta 1 in the renal tissue arranged in descending order were listed as follows: model group > treatment group > control group, and there were significant differences among three groups (P < 0.05). In the model group, diffuse glomerular hyperplasia was observed with the presence of increased extracellular matrix, partial glomerular sclerosis, and interstitial infiltration of inflammatory cells; in the treatment group, glomerular hyperplasia, mesangial proliferation and inflammatory cell infiltration were all mitigated compared with the model group. Therefore, bone marrow mesenchymal stem cell transplantation may contribute to renal repair after mesangial proliferative glomerulonephritis, by inhibiting overexpression of transforming growth factor beta 1 in the kidney.
    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 

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    Role and mechanism of gastric cancer stem cells in gastric cancer invasion, metastasis and angiogenesis
    Huang Hui, Pan Zhi-jian
    2016, 20 (14):  2021-2026.  doi: 10.3969/j.issn.2095-4344.2016.14.006
    Abstract ( 419 )   PDF (3604KB) ( 382 )   Save

    BACKGROUND: Although tumor stem cells and their differentiated vascular cells, which are not sensitive to chemotherapy and molecular targeted therapy, reduce overall blood supply of the tumor, these cells facilitate the invasion and metastasis of the tumor, eventually resulting in treatment failure.

    OBJECTIVE: To investigate the effect of gastric cancer stem cells in stomach cancer invasion and metastasis as well as its mechanism in angiogenesis.
    METHODS: A total of 20 gastric cancer stem cells and 20 normal gastric cancer cells SGC7901 were cultured in the medium with DMEM/F12 (1:1), 2% B27 and 20 μg/L epidermal growth factors or cultured in 10% PBS, respectively. Invasion of gastric cancer stem cells was detected by sphere formation assay, colony formation assay and Transwell invasion assay. Levels of mRNA and protein expression of CD44 and E-cadherin were measured to observe tumor metastasis. Besides, angiogenesis was observed by cell scratch test, Transwell migration assay and ring test.
    RESULTS AND CONCLUSION: Gastric cancer cells adhered to the medium wall, and gastric cancer stem cells suspended in the medium, which appeared to be spindle interstitial form. Number of gastric cancer tumor spheres was significantly lower than that of gastric cancer stem cell spheres in the sphere formation assay and colony formation assay (P < 0.05). Number of gastric cancer stem cells through the basement membrane was significantly higher than that of gastric cancer cells (P < 0.05). And the cell migration distance and ring number were significantly increased in the gastric cancer stem cells as compared with the gastric cancer cells (P < 0.05). In conclusion, the invasion, migration and angiogenesis abilities of gastric cancer stem cells are stronger than those of gastric cancer cells, which can further promote the occurrence and progression of gastric cancer. 
    中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 
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    Proliferative activity of bone marrow mesenchymal stem cells cultured in vitro: assessment by a CT brain perfusion scan
    Fu Feng-kui, Zhou Hai, Wang Jun
    2016, 20 (14):  2027-2032.  doi: 10.3969/j.issn.2095-4344.2016.14.007
    Abstract ( 187 )   PDF (1031KB) ( 317 )   Save

    BACKGROUND: Bone marrow mesenchymal stem cell transplantation is a hot spot in the treatment of ischemic cerebrovascular diseases, and CT brain perfusion scan is mostly used to observe the improvement in the blood flow in the ischemic region. However, X-ray irradiation during CT brain perfusion may affect the viability of transplanted bone marrow mesenchymal stem cells, and then influence the therapeutic efficacy.

    OBJECTIVE: To investigate the effect of CT brain perfusion dose on the viability of bone marrow mesenchymal stem cells.
    METHODS: Passage 5 bone marrow mesenchymal stem cells from Sprague-Dawley rats were selected and randomized into three groups, with 6 tubes in each group. One of the six tubes in each group was randomly selected with no irradiation, and the remaining five tubes in each group were subjected to CT brain perfusion scans 1-5 times, respectively. After scanning, the cell number was counted after 10-day continuous culture and cell growth curve of each tube was drawn. Cell cycle was detected by flow cytometry, and cell viability was measured by MTT method.
    RESULTS AND CONCLUSION: After 10 days of continuous counting, the number of cells per tube showed no difference (P > 0.05), and cell growth curves were basically coincided. Moreover, there was no significant difference in the cell cycle and cell viability (P > 0.05). Experimental findings show that the CT brain perfusion scan has no effect on the viability 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 
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    Cytobine-induced killer cells promote apoptosis of human liver cancer stem cells
    Shan Hai-xia, Fan Chong-gui, Huo Li-ya, Zhang Huai-hong, Zhai Yu-feng
    2016, 20 (14):  2033-2039.  doi: 10.3969/j.issn.2095-4344.2016.14.008
    Abstract ( 251 )   PDF (1288KB) ( 451 )   Save

    BACKGROUND: Immunotherapy with autologous immune cells has been developed as a major adjuvant therapy for malignant tumors, but its mechanism of action has not been elucidated.
    OBJECTIVE: To investigate the relationship between cytokine-induced killer cell secretion and apoptosis in human liver cancer stem cells.
    METHODS: Human liver cancer stem cells, HepG2 cells, were isolated and enriched using serum-free suspension method. The peripheral blood mononuclear cells from patients with liver cancer were induced by γ-interferon, CD3 monoclonal antibody and recombinant human interleukin-2 to form killer cells. Passage 1 liver cancer stem cells were divided into control group (culture alone) and experimental group (co-culture of cytokines-induced killer cells and human liver cancer stem cells). At 48 hours after culture, apoptosis in human liver cancer stem cells was detected using flow cytometry, and expression of caspase-3 mRNA and protein was detected using RT-PCR and western blot, respectively.
    RESULTS AND CONCLUSION: The apoptotic rate in the control group was significantly lower than that in the experimental group (P < 0.05). The expressions of caspase-3 at mRNA and protein levels were both higher in the experimental group than the control group (P < 0.05). Experimental findings show that cytokines-induced killer cells can significantly promote apoptosis in human liver cancer stem cells, and up-regulate the caspase-3 mRNA and protein expressions dramatically. 
    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 

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    Morphology and proliferation of bone marrow mesenchymal stem cells and expressions of CD34 and CD44 under stomach cancer microenvironment
    Wu Gao-feng, Liu Xi-ping, Yang Bai-lin, Li Pei-qing, Ming Hai-xia, Zhang Wei
    2016, 20 (14):  2040-2045.  doi: 10.3969/j.issn.2095-4344.2016.14.009
    Abstract ( 382 )   PDF (1208KB) ( 344 )   Save

    BACKGROUND: Gastric cancer mesenchyal stem cells from clinical stomach cancer specimens and tumorigenic tissues in nude mice are similar to the bone marrow mesenchymal stem cells in biological characteristics, which have been proved to be an important component of tumor microenvironment to promote tumor growth. It is speculated that biological characteristic of bone marrow mesenchymal stem cells may change in stomach cancer microenvironment.

    OBJECTIVE: To observe the effect of stomach cancer microenvironment on morphology and proliferation of bone marrow mesenchymal stem cells and expressions of CD34 and CD44.
    METHODS: Rat bone marrow mesenchymal stem cells were cultured alone as control group. In the test group, rat bone marrow mesenchymal stem cells were co-cultured with human stomach cancer BGC-823 cells using Transwell chamber assay to establish the stomach cancer microenvironment. Then, cell morphology, proliferation, cell cycle and CD34, CD44 expressions were observed and detected using inverted phase contrast microscope, MTT assay, and flow cytometry, respectively.
    RESULTS AND CONCLUSION: In the test group, bone marrow mesenchymal stem cells were similar to human stomach cancer cells BGC-823 that arranged disorderly and irregularly, were interconnected loosely, became thinner and longer, and grew in clusters with smaller nuclei. The cell proportion in G1 phase significantly decreased, but that in S and G2/M phases significantly increased (P < 0.01, P < 0.05). The positive rate of CD44 significantly declined, and the CD34 expression significantly raised (P < 0.01). In conclusion, stomach cancer microenvironment by non-contact co-culture with BCG-823 cells has an obvious effect on the morphology, proliferation and surface antigens expressions of bone marrow mesenchymal stem cells that will tend to be malignant gastric cancer 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 
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    Effects of bone marrow mesenchymal stem cells on the biological behavior of prostate cancer cells
    Liu Jian-jun, Duan Xiao-yu
    2016, 20 (14):  2046-2051.  doi: 10.3969/j.issn.2095-4344.2016.14.010
    Abstract ( 288 )   PDF (893KB) ( 343 )   Save

    BACKGROUND: Bone marrow mesenchymal stem cells have a close relationship with tumor occurrence and development, which are not only involved in tumor recruitment but impact the biological behavior of tumor cells.

    OBJECTIVE: To investigate the effect of bone marrow mesenchymal stem cells on the biological behavior prostate cancer cell line.
    METHODS: Human bone marrow mesenchymal stem cells from healthy adults were collected and cultured in vitro. Cell morphology and cell markers were observed. Using the Transwell technique, human dermal fibroblasts HDF-a were co-cultured with human prostate carcinoma DU145 cells in positive control group, wild-type prostate carcinoma DU145 cells cultured alone in negative control group, and bone marrow mesenchymal stem cells co-cultured with human prostate carcinoma DU145 cells in experimental group. Flow cytometry assay was used to detect the cell cycle of prostate carcinoma DU145 cells, soft agar colony formation assay performed to detect cell proliferative ability, and crystal violet staining used to observe DU145 cell migration at 3 days after culture.
    RESULTS AND CONCLUSION: Compared with the negative control group, cell proportion at G2/M+S phase, number of transmembrane cells and colony counting were significantly decreased in the experimental group. These findings indicate that bone marrow mesenchymal stem cells may inhibit the proliferation and invasion of prostate carcinoma DU145 cells by shortening the G2/M+S phase, thereby providing a new insight into the treatment of human prostate cancer.
    中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 
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    Emodin effects on renal ischemia/reperfusion injury after bone marrow mesenchymal stem cell transplantation
    Li Zhong
    2016, 20 (14):  2052-2058.  doi: 10.3969/j.issn.2095-4344.2016.14.011
    Abstract ( 233 )   PDF (1608KB) ( 320 )   Save

    BACKGROUND: Studies have shown that emodin protects against intestinal ischemia-reperfusion injury by inhibiting the release of inflammatory factors.

    OBJECTIVE: To investigate the effect of emodin on renal ischemia/reperfusion injury after bone marrow mesenchymal stem cell transplantation.
    METHODS: Forty Sprague-Dawley rats were randomized into five groups (n=8 per group): sham group, model group (renal ischemia/reperfusion injury group), bone marrow mesenchymal stem cell group (cell transplantation group), low-dose emodin group, high-dose emodin group. Rats in the latter four groups were pretreated with or without different concentrations (30 and 60 mg/kg) of emodin for 7 days, and then were subjected to clamping bilateral renal pedicles for 45 minutes, followed by injection of 1 mL bone marrow mesenchymal stem cell suspension. Six hours after reperfusion, the pathological changes of renal tissues were examined by hematoxylin-eosin staining; the mRNA levels of tumor necrosis factor α, interleukin-6, interleukin-18, TLR2 and TLR4 detected by real-time fluorescence quantitative PCR; and the protein expression of COX-2, ICAM-1 and iNOS determined by western blot.
    RESULTS AND CONCLUSION: Compared with the sham group, rats in the model group showed obvious features of severe acute tubular damage and inflammatory cell infiltration. In the cell transplantation group, tubular epitheliael cells were partially lost with some inflammatory cells infiltrated in the renal interstitium. In the emodin groups, the tubular lumen was practically intact with little renal interstitial inflammatory cells. Compared with the sham group, a significant increase in the mRNA levels of tumor necrosis factor α, interleukin-6, interleukin-18, TLR2 and TLR4 as well as in the protein levels of COX-2, ICAM-1 and iNOS was found in the model group (both P < 0.05). However, bone marrow mesenchymal stem cell transplantation attenuated this ischemia/reperfusion-induced increase in the expression of the above-mentioned factors (P < 0.05). Furthermore, the effects of bone marrow mesenchymal stem cell transplantation were further augmented by pretreatment with emodin in a dose-dependent manner. These findings suggest that emodin can enhance the protective effects of bone marrow mesenchymal stem cells on renal ischemia/reperfusion 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 
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    Bone marrow mesenchymal stem cells transplanted into a rat model of Alzheimer’s disease: improvement in the learning and memory ability
    Gao Ming-long, Zhang Ying-dong, Li Na, Qiao Jun, Yu Ming
    2016, 20 (14):  2059-2065.  doi: 10.3969/j.issn.2095-4344.2016.14.012
    Abstract ( 342 )   PDF (2118KB) ( 310 )   Save

    BACKGROUND: Drug therapy can partly reduce and delay the progress of Alzheimer’s disease, but only 30% with the single drug treatment obtain clinical cure.
    OBJECTIVE: To study the therapeutic effect of bone marrow mesenchymal stem cells transplantation for rats with Alzheimer’s disease.
    METHODS: Amyloid β-protein was injected into the hippocampus of Sprague-Dawley rats to construct the model of Alzheimer’s disease. And bone marrow stromal stem cells were transplanted into the hippocampus of the rat models.
    RESULTS AND CONCLUSION: At 2 weeks after modeling, compared with the control group, the escape latency in the model and experimental groups was significantly longer (P < 0.05), which indicating that Alzheimer’s disease models were successfully established. At 4 weeks after cell transplantation, compared with the model group, the average escape latency in the experimental group was significantly decreased, but retention time on the platform quadrant was significantly prolonged (P < 0.05). Besides, at 4 weeks after cell transplantation, expression of choline acetyltransferase in the experimental group was significantly higher than that in the model group (P < 0.05). In conclusion, bone marrow mesenchymal stem cells cannot only differentiate and survive in the hippocampus of rats with Alzheimer’s disease, but also improve 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

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    CD133+ cells in combination with human umbilical cord stem cells in mouse heart failure
    Hou Mei, Zhang Hong-xing, Ye Qing, Zhang Yong
    2016, 20 (14):  2066-2072.  doi: 10.3969/j.issn.2095-4344.2016.14.013
    Abstract ( 298 )   PDF (4254KB) ( 314 )   Save

    BACKGROUND: Currently, conventional treatment methods for heart failure are all ineffective.
    OBJECTIVE: To explore the combined effects of human umbilical cord stem cells and CD133+ cells in mice with heart failure, providing a new insight into the treatment of heart failure.
    METHODS: Full-term newborn umbilical cord from vaginal delivery was collected to isolate CD133+ cells and human umbilical cord stem cells using lymphocyte separation medium method. Twenty Balb/C nude mice were randomly subjected to mononuclear cell injection (mononuclear cell group) or injection of CD133+ cells combined with human umbilical cord stem cells (combined group) via the tail vein after establishing heart failure models.
    RESULTS AND CONCLUSION: Fourteen days after injection, the body weight and liver, heart and lung mass of mice were significantly larger in the combined group than the mononuclear cell group (P < 0.05). After 30 days, myocardial cells arranged regularly in the combined group, but disorderly in the mononuclear cell group; compared with the mononuclear cell group, the average area of myocardial collagen fibers was significantly decreased in the combined group (P < 0.05), and the level of serum matrix metalloproteinase-9 was also significantly lower in the combined group (P < 0.05). Masson staining showed that blue-stained collagen fibers in the combined group were less but arranged neatly; however, in the mononuclear cell group, the number of collagen fibers that arranged irregularly was increased to different extents. To conclude, the combined use of CD133+ cells and human umbilical cord stem cells has desired outcomes in the treatment of heart failure in mice, indicating a higher clinical value.
    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

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    Bone marrow mesenchymal stem cell transplantation for gastric precancerous lesions in a rat model
    Liu Hong-feng, Li Lu
    2016, 20 (14):  2073-2079.  doi: 10.3969/j.issn.2095-4344.2016.14.014
    Abstract ( 377 )   PDF (1320KB) ( 222 )   Save

    BACKGROUND: Precancerous lesions are a long-term development process in which many factors are involved. Bone marrow mesenchymal stem cells can repair tissue injury.

    OBJECTIVE: To investigate the effect of bone marrow mesenchymal stem cell transplantation on gastric precancerous lesions in the rats.
    METHODS: Thirty-six Wistar rats were randomly divided into control group, model group and transplantation group. Animal models of gastric precancerous lesions were established in the model and transplantation groups. Rats in the transplantation group were given 1 mL of CM-dil-labeled bone marrow mesenchymal stem cells (3×106 cells) via the tail vein, once a week, totally three times. Rats in the model and control group were subjected to the tail vein injection of the same volume of normal saline. Then, rats were sacrificed 1 week after final injection, and pathohistological changes in rat gastric tissue sections were observed. The expression of vascular endothelial growth factor in the gastric mucosa and levels of serum cytokines were detected.
    RESULTS AND CONCLUSION: The severity of gastric mucosal injury in the transplantation group was lighter than that of the model group. The expression of vascular endothelial growth factor was significantly higher in the transplantation group compared with the model and control groups (P < 0.05). The levels of serum interleukin-17 and interferon-γ were significantly higher in the model group than the transplantation group followed by the control group (both P < 0.05). Therefore, bone marrow mesenchymal stem cells can increase vascular permeability, reduce inflammation, block or ease the occurrence of precancerous lesions by up-regulating the expression of vascular endothelial growth factor in the gastric mucosa lesions and reducing the expression of interleukin-17 and interferon-γ.
    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
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    Umbilical cord blood stem cell transplantation for treatment of decompensated cirrhosis
    Guo Xian-li, Liu Yue, Zhou Li-min, Hu Yu
    2016, 20 (14):  2080-2085.  doi: 10.3969/j.issn.2095-4344.2016.14.015
    Abstract ( 380 )   PDF (1229KB) ( 304 )   Save

    BACKGROUND: Umbilical cord blood stem cells can be differentiated into functional liver cells in vivo, instead of mature liver cells in the treatment of liver diseases.

    OBJECTIVE: To observe the effect of umbilical cord blood stem cell transplantation on decompensated cirrhosis in rats.
    METHODS: Twenty Sprague-Dawley rats were enrolled and given 40% carbon tetrachloride peanut oil to make decompensated cirrhosis models. Eight weeks after modeling, model rats were randomized into control and experimental groups (n=10 per group), and subjected to injection of umbilical cord blood stem cell suspension (2 mL, 1.0×107/L) or cell culture medium (2 mL) via the tail vein, respectively. At 4 weeks after injection, levels of serum aspartate aminotransferase, alanine aminotransferase, albumin, total bilirubin, hyaluronic acid, laminin, procollagen III and type IV collagen were detected; rat liver tissues were sliced for pathological observation.
    RESULTS AND CONCLUSION: Compared with the control group, the albumin level was significantly increased but the levels of serum aspartate aminotransferase, alanine aminotransferase, total bilirubin, hyaluronic acid, laminin, procollagen III and type IV collagen decreased significantly in the experimental group (P < 0.05). From the pathological view, severity of liver cirrhosis was lower in the experimental group than the control group. Therefore, umbilical cord blood stem cell transplantation can effectively restore the liver function of decompensated cirrhosis in 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
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    Autologous peripheral blood stem cell transplantation combined with percutaneous transluminal angioplasty for diabetic lower extremity vascular disease
    Gu Lu, Zhang Shu-mei, Yu Xiang, Yang Yi
    2016, 20 (14):  2086-2091.  doi: 10.3969/j.issn.2095-4344.2016.14.016
    Abstract ( 402 )   PDF (1041KB) ( 531 )   Save

    BACKGROUND: Studies have found that peripheral blood stem cells can highly differentiate into vascular endothelial cells to promote blood vessel regeneration, and improve collateral circulation, thereby achieving satisfactory outcomes in the treatment of limb ischemia.

    OBJECTIVE: To observe the clinical effect in diabetic lower extremity vascular disease patients undergoing percutaneous transluminal angioplasty combined with autologous peripheral blood stem cell transplantation.
    METHODS: Fifty patients hospitalized for diabetic lower extremity vascular disease from March in 2011 to December in 2014 were collected: 25 cases underwent percutaneous transluminal angioplasty as control group; another 25 underwent percutaneous transluminal angioplasty combined with autologous peripheral blood stem cell transplantation as combination group. At 1, 6, 12 months after surgery, subjective scores of affected limb pain and cold sensation were recorded, additionally, objective indicators, including ankle-brachial index, transcutaneous oxygen partial pressure, and claudication distance were detected.
    RESULTS AND CONCLUSION: Ankle-brachial index scores of the two groups were significantly increased, especially at 1 month after surgery, but decreased at 6, 12 months, and there was a significant difference compared with that before treatment (P < 0.05); at 1, 6, and 12 months, there was no significant difference between the two groups (P > 0.05). At 1, 6, and 12 months after surgery, both of transcutaneous oxygen partial pressure and claudication distance in the two groups were significantly increased compared with those before treatment (P < 0.05). Furthermore, compared with the control group, these two indicators in the combination group were significantly increased (P < 0.05). Within 12-month follow-up, affected limb pain and cold sensation were improved in the two groups, especially in the combination group. Inevitably, three cases had hypocalcemia during the collection of peripheral blood stem cells, and two cases developed fever and 3 cases appeared to have local exudation after surgery. All these symptoms were released by symptomatic treatments, respectively. In conclusion, percutaneous transluminal angioplasty combined with autologous peripheral blood stem cell transplantation for diabetic lower extremity vascular lesions can promote the establishment of affected limb collateral circulation, to decrease the risk of limb ischemia, which achieves significant outcomes than percutaneous transluminal angioplasty used alone.
    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
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    Neural stem cell transplantation for treatment of cerebral palsy
    Lou Yuan-jun, Liu Yang, Shan Hai-jun, Cao Cai-hong, Jie Xiao-su
    2016, 20 (14):  2092-2097.  doi: 10.3969/j.issn.2095-4344.2016.14.017
    Abstract ( 284 )   PDF (1043KB) ( 261 )   Save

    BACKGROUND: Transplanted neural stem cells can survive, proliferate and differentiate into neurons and/or glial cells in the host, thereby promoting partial function recovery in the host.

    OBJECTIVE: To study the therapeutic effects of neural stem cell transplantation on cerebral palsy rats.
    METHODS: Twenty-four rats were randomly divided into three groups: control group, model group and transplantation group. Animal models of cerebral palsy were made in the latter two groups. One week after modeling, rats in the transplantation group were injected 1 mL stem cell suspension (1×105) via the jugular vein, and rats in the control and model group were given the same volume of normal saline. Toe distance, step length and elevated body swing test in rats were detected, and histopathological changes in the rat brain were observed 3 weeks after transplantation.
    RESULTS AND CONCLUSION: In the model group, the toe distance and step length of the front left palm were significantly lower than those of the front right palm (P < 0.05). Compared with the model group, the toe distance and step length in the transplantation and control groups were significantly increased (P < 0.05). In the elevated body swing test, rats in the model group presented with asymmetric swing of the body, but rats in the other two groups exhibited symmetric swing of the body (P < 0.05). Additionally, the ratio of right to left hemispheric areas was significantly higher in the transplantation and control groups compared with the model group (P < 0.05). In conclusion, neural stem cell transplantation via the jugular vein can improve brain function and restore motor function in rats with cerebral palsy.
    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
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    Endometrial stromal stem cells from early-pregnancy decidua differentiate into endometrial epithelial cells
    Chen Bo
    2016, 20 (14):  2098-2103.  doi: 10.3969/j.issn.2095-4344.2016.14.018
    Abstract ( 385 )   PDF (1057KB) ( 366 )   Save

    BACKGROUND: Decidua tissue is one of the most important sources of endometrial stem cells, from which stromal stem cells can differentiate into various kinds of cells under contain inductions.

    OBJECTIVE: To observe the differentiation of endometrial stromal stem cells isolated from early-pregnancy decidua into endometrial epithelial cells.
    METHODS: Endometrial stromal stem cells from early-pregnancy decidua were cultured, identified and directionally induced. Then induced cells were randomized into test group cultured in conditioned medium and control group cultured in normal culture medium containing fetal bovine serum. Expression of keratin was detected by immunofluorescence assay and western bolt assay in the two groups.
    RESULTS AND CONCLUSION: The endometrial stromal stem cells from early-pregnancy decidua were successfully isolated and cultured, and presented with a S-shaped curve. Within 1-48 hours after inoculation, cultured cells were in the adaptation phase; then, the cells proliferated rapidly and began to grow logarithmically at 48 hours after culture; after 60 hours of culture, the cell growth rate decreased, with a steady rise in the cell curve. Results from the flow cytometry detection showed that the endometrial stromal stem cells could express CD73 and CD90, but not express CD34, CD45 and cytokeratin. Immunofluorescence findings showed the expression of keratin negative in the control group, and mostly positive in the test group. Besides, the relative expression of keratin in the test group was significantly higher than that in the control group (P < 0.05). These results suggest that the endometrial stromal stem cells can be isolated from the human decidua in early pregnancy, and differentiate into endometrial epithelial cells by directional induction.
    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
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    Seminiferous capsule extract effect on the proliferation of spermatogonial stem cells
    Li Jun-tao, Zhang Pei-hai, Qu Xiao-wei, Li Zheng-sheng, Li Song-wei
    2016, 20 (14):  2104-2109.  doi: 10.3969/j.issn.2095-4344.2016.14.019
    Abstract ( 301 )   PDF (1072KB) ( 255 )   Save

    BACKGROUND: Chinese herbs for kidney nourishment can promote the proliferation and differentiation of spermatogonial stem cells.

    OBJECTIVE: To verify the effect of seminiferous capsule extract on spermatogonial stem cell proliferation.
    METHODS: Spermatogonial stem cells were isolated from the testis of male mice and synchronized by serum-free medium followed by an addition of 10, 50, 100 mg/L seminiferous capsule extracts. After 24 hours of culture, viability, proliferation and cell cycle of spermatogonial stem cells were observed.
    RESULTS AND CONCLUSION: Compared with the control group, seminiferous capsule extracts promoted the cell number, viability and proportion at S stage. The number of BrdU-labeled spermatogonial stem cells was increased significantly after intervention with seminiferous capsule extracts, especially at the concentration of 50 mg/L. These findings indicate that seminiferous capsule extracts can promote the proliferation and viability of spermatogonial 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
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    Electrotransfection by human telomerase reverse transcriptase gene: optimization in precartilagious stem cell culture
    Peng Yi, Qu Jia-fu, Cao Li-hai, Zhao Guo-zhi, Du Xiao-jian
    2016, 20 (14):  2110-2116.  doi: 10.3969/j.issn.2095-4344.2016.14.020
    Abstract ( 251 )   PDF (1191KB) ( 281 )   Save

    BACKGROUND: The human telomerase reverse transcriptase gene (hTERT) transfected into target cells can play an important role in target cell proliferation and differentiation by increasing telomerase activity and maintaining telomere length.

    OBJECTIVE: To explore the effect of hTERT transfection on telomerase activity and biological characteristics of precartilage stem cells culured in vitro.
    METHODS: Precartilage stem cells cultured in vitro were subjected to hTERT gene transfection via a retrovirus vector pLXSN. Meanwhile, control and negative control groups were set up. After transfection, TRAP-ELISA assay was used to detect telomerase activity; RT-PCR and western blot employed to detect hTERT mRNA and protein expressions; cell counting kit-8 used to detect cell proliferaiton based on cell growth curve; and flow cytometry adopted to detect cell cycle and distribution.
    RESULTS AND CONCLUSION: The telomerase activity was significantly increased at 48 hours after hTERT gene was transfected into the precartilage stem cells. After transfection of hTERT, hTERT mRNA and protein levels were significantly increased, the cell growth rate was significantly increased, the proportion of cells at G0/G1 phase was decreased, and the number of S-phased cells increased compared with the control group and negative control group. There were significant differences among the groups (P < 0.05). In conclusion, hTERT transfection via retrovirus vector pLXSN can promote the proliferation of precartilage stem cells in rats by increasing the telomerase activity.
    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
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    Human umbilical cord mesenchymal stem cells induced by 5-azacytidine: bioactive properties and cardiomyocyte-like differentiation
    Li Ping, Huangpu Chun-rong, Tang Chun-hui, Chen Yi-hui
    2016, 20 (14):  2117-2122.  doi: 10.3969/j.issn.2095-4344.2016.14.021
    Abstract ( 313 )   PDF (3570KB) ( 432 )   Save

    BACKGROUND: Human umbilical cord mesenchymal stem cells belong to a special class of stem cells with a limited number, which are difficult to isolate and purify. Importantly, there is a lack of clinical understanding of biological characteristics of human umbilical cord mesenchymal stem cells.

    OBJECTIVE: To study the bioactive properties and cardiomyocyte-like differentiation of human umbilical cord mesenchymal stem cells.
    METHODS: Under a sterile collection, umbilical cord blood of newborn from cesarean section was collected to isolate human mesenchymal stem cells by digestion using collagenase II. Immunohistochemical staining was used to detect cell surface markers and observe morphological characteristics, and MTT assay used to detect cell proliferation and draw cell growth curve. Cells grown to 70%-80% confluence were treated with 10 μmol/L 5-azacytidine for 24 hours.
    RESULTS AND CONCLUSION: One week after adherent tissue culture, human umbilical cord mesenchymal stem cells were seen, and then the cells cultured in culture medium showed colony growth. Within 24 hours of cell seeding, cells at passages 3 and 10 were at latency period and then grew logarithmically with a doubling time of 30 hours. Immunocytochemical staining showed that passage 3 cells at over 90% confluence expressed CD44 and CD29, rather than CD28 and CD31.Three weeks after induced culture, the cells with the absence of processes grew irregularly; after 4 weeks, the cells were round in shape and expressed cardiac-specific immune markers. Taken together, isolated human umbilical cord mesenchymal stem cells with strong proliferative ability have the basic biological characteristics of mesenchymal stem cells, which are capable of differentiating into cardiomyocyte-like cells induced by 5-azacytidine.
    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
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    Effect of umbilical cord blood mesenchymal stem cell transplantation on myocardial cell apoptosis
    Wang Yan-li, Li Jin-feng, Wang Yan-ping, Gao Ming
    2016, 20 (14):  2123-2128.  doi: 10.3969/j.issn.2095-4344.2016.14.022
    Abstract ( 380 )   PDF (4234KB) ( 291 )   Save

    BACKGROUND: Umbilical cord blood mesenchymal stem cell transplantation can reduce myocardial apoptosis and myocardial fibrosis, thereby improving the cardiac function.

    OBJECTIVE: To observe the effect of umbilical cord blood mesenchymal stem cell transplantation on myocardial apoptosis in rats.
    METHODS: (1) In vitro test: Normal myocardial cells cultured for 72 hours were enrolled as control group. Myocardial cells injured by adriamycin alone were enrolled as injury group. Adriamycin-injured myocardial cells were co-cultured with umbilical cord blood mesenchymal stem cells as co-culture group. After 48 hours of culture, TUNEL assay was used to assess myocardial apoptosis in rats. (2) In vivo test: Forty-five Sprague-Dawley rats were randomized into normal control, adriamycin, and umbilical cord blood mesenchymal stem cell transplantation (cell transplantation) groups. Two weeks after cell transplantation, PowerLab was used to test cardiac function, and western blot employed to detect Bax and Caspase-3 expression levels. Meanwhile, pathological changes of the myocardial tissues were observed in the three groups.
    RESULTS AND CONCLUSION: (1) In vitro test: The apoptotic rate of myocardial cells was significantly increased in the injury group compared with the control group, but decreased significantly in the co-culture group compared with the injury group (P < 0.05). (2) In vivo test: In the cell transplantation group, the cardiac function indexes, expression levels of Bax and Caspase-3 and myocardial infarction size were significantly improved compared with the adriamycin group (P < 0.05), but still not recovered to the normal levels (P < 0.05). These results show that umbilical cord blood mesenchymal stem cells acting on the rat myocardial cells can inhibit adriamycin-induced myocardial apoptosis, and the relevant mechanism is probably related to down-regulation of Bax and caspase-3 expression.
    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
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