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    06 May 2016, Volume 20 Issue 19 Previous Issue    Next Issue
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    Human preproenkephalin gene-modified bone marrow mesenchymal stem cell line: its biological characteristics
    Li Xin-mao
    2016, 20 (19):  2741-2747.  doi: 10.3969/j.issn.2095-4344.2016.19.001
    Abstract ( 293 )   PDF (961KB) ( 414 )   Save

    BACKGROUND: Previous studies have found that the transplantation of modified cell lines exhibit analgesic effect. But little is reported on the biological characteristics of human preproenkephalin gene-modified bone marrow mesenchymal stem cell lines.
    OBJECTIVE: To observe the biological characteristics of human preproenkephalin gene-modified bone marrow mesenchymal stem cells.
    METHODS: Bone marrow mesenchymal stem cells were isolated and cultured to establish human preproenkephalin gene-modified bone marrow mesenchymal stem cell lines.
    RESULTS AND CONCLUSION: After freezing-thawing, passage 4 human bone marrow mesenchymal stem cells, human bone marrow mesenchymal stem cells-pBABE and human bone marrow mesenchymal stem cells-human preproenkephalin exhibited no significant changes in the cell viability (P > 0.05), as well as in the fat cell proportion after adipogenic induction (P > 0.05). Moreover, red calcium deposition was presented in all these cells by alizarin red staining after osteogenic induction. Flow cytometry results showed that passage 4 human bone marrow mesenchymal stem cells, human bone marrow mesenchymal stem cells-pBABE and human bone marrow mesenchymal stem cells-human preproenkephalin could express CD29 and CD44, but not express CD34 and CD45. Human preproenkephalin genes were highly expressed in human bone marrow mesenchymal stem cells-pBABE, but lowly expressed in passage 4 human bone marrow mesenchymal stem cells. Additionally, recombinant plasmid pBABE-preproenkephalin-modified human bone marrow mesenchymal stem cells could express enkephalin protein. In conclusion, human preproenkephalin gene-modified human bone marrow mesenchymal stem cell lines can maintain the pluripotent differentiation or proliferation capacity of bone marrow mesenchymal stem cells, and secrete enkephalin protein.

     

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    Bone marrow mesenchymal stem cells combined with endothelial progenitor cells for repair of alveolar bone defect in ovariectomized rats
    Wen Yi, Yang Hong-xu, Liu Qian, Liang Yuan, Liu Zhu-ying, Ding Yin
    2016, 20 (19):  2748-2755.  doi: 10.3969/j.issn.2095-4344.2016.19.002
    Abstract ( 269 )   PDF (2296KB) ( 428 )   Save

    BACKGROUND: Numerous studies have demonstrated that estrogen can regulate the proliferation and migration of endothelial progenitor cells (EPCs), while EPCs can also promote the function and activity of bone marrow mesenchymal stem cells (BMSCs) in vitro.
    OBJECTIVE: To evaluate the ability of the BMSCs and EPCs which construct the composite cell sheet in the repair of alveolar bone defect in ovariectomized rats.
    METHODS: BMSCs/EPCs composite sheet, EPCs sheet and BMSCs sheet were respectively implanted into the defects of the alveolar bone in ovariectomized rats. Rats with no implantation served as control group. Repaired alveolar bone was assessed by gross examination, histological observation and micro-CT scan at 2, 4, 8 weeks after operation.
    RESULTS AND CONCLUSION: BMSCs/EPCs composite sheet has greater osteogensis activity and bone repair capacity than BMSCs or EPCs sheet alone.

     

     

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    Bone marrow mesenchymal stem cells co-cultured with allogenic bone in the articular cavity
    Gao Feng-guang, Zhou Bai-sui, Mu Hai-bo
    2016, 20 (19):  2756-2762.  doi: 10.3969/j.issn.2095-4344.2016.19.003
    Abstract ( 280 )   PDF (1962KB) ( 333 )   Save

    BACKGROUND: Bone marrow mesenchymal stem cells and allogenic bones are commonly used as seed cells and scaffolds, respectively, for constructing tissue-engineered cartilage through in vitro co-culture. The loose body of the knee joint can survive in the articular cavity for a long time, and maintain certain characteristics of cartilage tissues. Therefore, the articular cavity can provide a good environment for the growth and development of chondrocytes.
    OBJECTIVE: To investigate the effect of bone marrow mesenchymal stem cells co-cultured with allogenic bone in the articular cavity.
    METHODS: Bone marrow mesenchymal stem cells were isolated from five newborn New Zealand white rabbits. One adult New Zealand rabbit was enrolled to prepare allogenic bone for co-culture with bone marrow mesenchymal stem cells. Afterwards, bone marrow mesenchymal stem cells and allogenic bone composites were cultured in the articular cavity (intracavitary culture group) or in vitro as in vitro culture group, respectively; the normal cartilage tissues grew in the articular cavity as control group. Cells were observed by hematoxylin-eosin staining and type II collagen immunohistochemistry staining at 4, 8 and12 weeks of culture.
    RESULTS AND CONCLUSION: At 12 weeks culture, hematoxylin-eosin staining showed: in the control group, chondrocytes arranged tightly and directionally with red stained cytoplasm and cartilage matrix as well as blue nuclei; in the intracavitary culture group, the scaffold was mostly absorbed and chondrocytes grew into the scaffold in a certain direction with smaller shape, while cytoplasm and cartilage matrix were red stained, blue nuclei appeared; in the in vitro culture group, abundant chondrocytes proliferated in a disordered arrangement. Immunohistochemistry staining showed: the absorbance (A) values in the intracavitary culture group showed a continuous increase, but no obvious change was in the other two groups. Moreover, at 4, 8 and 12 weeks of culture, A values in the control group and intracavitary culture group were significantly higher than that in the in vitro culture group (P < 0.05); at 4 and 8 weeks, A value in the intracavitary culture group was significantly lower than that in the control group (P < 0.05), but at 12 weeks, there was no significant difference in A value between the two groups (P > 0.05). These results suggest that the tissue-engineered cartilage can be constructed by bone marrow mesenchymal stem cells co-cultured with allogenic bone under in vitro and in vivo environment, especially in the articular cavity.

     

     

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    Construction of Nanog lentiviral expression vector and its application in differentiation regulation of embryonic stem cells
    Chen Fei, Wan Li, Li Yan, Li Xin
    2016, 20 (19):  2763-2769.  doi: 10.3969/j.issn.2095-4344.2016.19.004
    Abstract ( 313 )   PDF (1674KB) ( 566 )   Save

    BACKGROUND: There is a lack of safe and effective modular lentivectors in differentiation regulation of embryonic stem cells.
    OBJECTIVE: To construct a modular lentivector, Puro-Nanog-hrGFP, with Nanog promoter-controlled humanized renilla reniformis green fluorescent protein (hrGFP) and puromycin and to observe the expression of Nanog during the differentiation of Nanog-hrGFP-transfected mouse embryonic stem cell lines.
    METHODS: After PCR amplification, Nanog, hrGFP and entry vector were recombined into the pDest-puro vectors to generate the lentiviral expression vector, Puro-Nanog-hrGFP, by the LR reaction. Through lentivirus production, transduction and puromycin screening, the transduced cell lines with Nanog-hrGFP gene were generated and identified. Expression of Nanog during the differentiation of transgenic mouse cell lines was detected.
    RESULTS AND CONCLUSION: The lentiviral expression vectors Puro-Nanog-hrGFP were constructed successfully by Gateway technology, and then the transducted cell lines were obtained by lentiviral infection. The expression of Nanog was gradually decreased during the process of transgenic cell lines differentiation, which provides a new tool for further investigation on regulation of stem cell differentiation.

     

     

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    Differentiation of bone marrow mesenchymal stem cells into endometrial epithelial cells in vitro
    Qu Li-hong, Yu Jia-qun
    2016, 20 (19):  2770-2776.  doi: 10.3969/j.issn.2095-4344.2016.19.005
    Abstract ( 403 )   PDF (903KB) ( 825 )   Save

    BACKGROUND: Previous studies have confirmed that as an exogenous origin of endometrial epithelial cells, bone marrow stem cells are involved in the functional reconstruction of the injured endometrium.
    OBJECTIVE: To discuss whether bone marrow mesenchymal stem cells can differentiate into endometrial epithelial cells in vitro.
    METHODS: Passage 2 bone marrow mesenchymal stem cells and endometrial stromal cells were collected and subjected to different treatments: bone marrow mesenchymal stem cells were cultured alone in the DMEM/F12 medium containing 2% fetal bovine serum or in the DMEM/F12 medium containing 2% fetal bovine serum, 10-7 mol/L β-estradiol and 10 μg/L epidermal growth factor as groups 1 and 2, respectively; bone marrow mesenchymal stem cells co-cultured with endometrial stromal cells by Transwell chamber were cultured in the DMEM/F12 medium containing 2% fetal bovine serum or in the DMEM/F12 medium containing 2% fetal bovine serum, 10-7 mol/L β-estradiol and 10 μg/L epidermal growth factor as groups 3 and 4, respectively. After 5 days culture, bone marrow mesenchymal stem cells at the bottom of the culture plate were collected to detect expressions of epithelial cell markers, such as CK7, CK18, CK1 and EMA by RT-PCR technology, and to determine keratin expression using immunofluorescence assay. Besides, expressions of these epithelial cell markers in the group 3 were detected using RT-PCR at 1, 3, 5 and 7 days of culture, respectively.
    RESULTS AND CONCLUSION: mRNA expression of CK7 showed a significant successive rise in the groups 1, 2, 3, 4 and it was highest in the group 4. mRNA expressions of CK18 and CKl9 in the later three groups had no significant differences, but all significantly higher than those in the group 1. Compared with the groups 1 and 3, mRNA expression of EMA in the groups 2, 4 were significantly higher, but no significant differences existed between groups. Expression of keratin was strongly positive in the group 4, weakly positive in the group 3 and negative in the groups 1, 2, respectively. Furthermore, with the increase of culture time, mRNA expression of CK7 exhibited a constant increase, which was significantly higher at 5 and 7 days than at 1 and 3 days; but there were no significant differences in expressions of CKl9, CKl8 and EMA at different time points. These results show that bone marrow mesenchymal stem cells can differentiate into endometrial epithelial cells in vitro under certain conditions. Moreover, it can remarkably promote the endometrial differentiation under the combined effects of some exogenous and endogenous factors.

     

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    Simvastatin effects on the expressions of specific osteogenic genes in bone marrow stromal stem cells
    Liu Hao, Zhang Yan, Liu Jia-yin, Liu Guang-yuan, Zhang Ke-zhong, Zhang Guo-bin, Xing Lei,Tian Fa-ming
    2016, 20 (19):  2777-2782.  doi: 10.3969/j.issn.2095-4344.2016.19.006
    Abstract ( 276 )   PDF (895KB) ( 346 )   Save

    BACKGROUND: Previous studies have demonstrated that simvastatin that can promote osteogenic differentiation of bone marrow stromal stem cells in vitro, is likely to be a new osteogenic drug. While it is still unknown whether there is time-dependent stimulation of simvastatin on the expressions of bone morphogenetic protein 2 and collagen type I.
    OBJECTIVE: To investigate the expressions of bone morphogenetic protein 2 and collagen type I in rat bone marrow stromal stem cells in vitro stimulated by simvastatin at different time points.
    METHODS: Passage 1 bone marrow stromal cells were divided into control and simvastatin group, followed by cultured in osteogenic differetiation medium with or uithout 10-7 mol/L simvastatin. After 7-day intervention, expression of alkaline phosphatase was detected in passage 3 cells. Passage 4 cells were divided and cultured as described above, and afterwards, RNA and proteins were extracted at 12 and 36 hours to detect the expressions of bone morphogenetic protein 2 and collagen type I using real-time PCR and western blot assay.
    RESULTS AND CONCLUSION: Both two groups could express alkaline phosphatase, while the rate of positive cells significantly increased in the simvastatin group compared with the control group (P < 0.05); at 12 and 36 hours after intervention, mRNA expressions of bone morphogenetic protein 2 and collagen type I in the simvastatin group were significantly higher than those in the control group (P < 0.05). Besides, western blot assay showed: at both 12 and 36 hours, simvastatin significantly enhanced the expression of bone morphometric protein 2, while the expression of collagen type I significantly increased at 12 hours (P < 0.05), but not at 36 hours. In conclusion, simvastatin can promote the expressions of bone morphometric protein 2 and collagen type I in rat bone marrow stromal cells, with more favorable outcomes after 12-hour treatment.

     

     

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    Mechanism of bone marrow mesenchymal stem cells in promoting lung cancer metastasis
    Zheng Tian-liang, Zhao Song, Guo Hai-zhou, Cui Guang-hui, Lin Da-wei
    2016, 20 (19):  2783-2788.  doi: 10.3969/j.issn.2095-4344.2016.19.007
    Abstract ( 324 )   PDF (1067KB) ( 734 )   Save

    BACKGROUND: So far the positive or negative effects of mesenchymal stem cells on tumor growth and metastasis are under discussion.
    OBJECTIVE: To explore the mechanism of bone marrow mesenchymal cells in promoting lung cancer metastasis.
    METHODS: Primary rat bone marrow mesenchymal stem cells were obtained by direct adherence method of the whole bone marrow, and differential adherence combined with digestion control method was performed to purify cells. Lung cancer cell lines were cultured, and the effects of bone marrow mesenchymal stem cells on the migration, invasion and metastasis of lung cancer cells were observed by scratch test, cell invasion and migration assays. Orthotopic lung cancer models were established in rats and bone marrow mesenchymal stem cells were seeded onto the left lung of rats. Then, pathological changes of lung tissues were observed at 14 days after transplannation.
    RESULTS AND CONCLUSION: After the scratch test, the migration rate of lung cancer cells became higher, and the scratches healed with time. And after cell transplantation, the number of migrated lung cancer cells increased, as well as the ability of lung cancer cells penetrating the Matrigel was strengthened. Besides, fibrous connective tissues could be found around the lung cancer tissues, and necrosis with distinct boundary and large tumor nuclei; the metastatic tissues showed obvious infiltration and necrosis with large tumor nuclei. These results suggest that bone marrow mesenchymal stem cells can promote the invasion, migration and metastasis of lung cancer cell lines.

     

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    Combination of acupuncture with placenta derived mesenchymal stem cell transplantation for cerebral palsy in a rat model
    Shi Hua, Zhang Pu, Guo Xin, Jie Xiao-su
    2016, 20 (19):  2789-2795.  doi: 10.3969/j.issn.2095-4344.2016.19.008
    Abstract ( 303 )   PDF (2028KB) ( 341 )   Save

    BACKGROUND: Both of acupuncture and mesenchymal stem cell transplantation have therapeutic effects on cerebral palsy.
    OBJECTIVE: To explore the therapentic effect of acupuncture combined with placenta derived mesenchymal stem cells on rat cerebral palsy.
    METHODS: Thirty-five Wistar rats were equivalently randomized into five groups, including control group, model group, acupuncture group, transplantation group and combination group. Rat models of cerebral palsy were prepared in all groups except the control group. At 7 days after modeling, in the transplantation group, 10 μL placenta derived mesenchymal stem cells were transplanted into the right cerebral cortex and striatum of rats; rats in theacupuncture group were given acupuncture at Shuigou (DU26), Dazhui (DU14), Baihui (DU20), Guanyuan (RN4) and Qihai (RN6) points; rats in the combination group given acupuncture and transplantation treatment as described above; the model and control groups received no treatment. Then, after 7-day intervention, the body mass, neurological function and pathological changes of rat brain tissues were measured and observed, respectively.  .
    RESULTS AND CONCLUSION: The order of the growth rate of body mass was as follows: control group > combination group > acupuncture and transplantation groups > model group, and all groups had significant differences except the two groups of acupuncture and transplantation (P < 0.05); findings from holding tract, foot error and water finding in maze tests showed the neurological function was best in the control group, followed by the combination group, as well as acupuncture and transplantation groups, and the worst in the model group. All groups had significant differences except acupuncture and transplantation groups (P < 0.05). Besides, in the model group, cells in the brain and hippocampus decreased, arranging disorderly with fuzzy structure, and inflammatory cells appeared; pathological changes in the other groups were in between control group and model group, Additionally, brain tissues in the combination group exlibited better pathological changes than those in the acupuncture and transplantation groups, which were similar with the normal brain tissues. To conclude, acupuncture, placenta derived mesenchymal stem cells transplantation, or their combination can improve neurological function in rats with cerebral palsy, especially the combination treatment.

     

     

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    Rosuvastatin combined with umbilical cord blood mesenchymal stem cell transplantation improves cardiac function after acute myocardial infarction
    Wang Yan-li, Li Jin-feng, Wang Yan-ping, Gao Ming
    2016, 20 (19):  2796-2802.  doi: 10.3969/j.issn.2095-4344.2016.19.009
    Abstract ( 379 )   PDF (1431KB) ( 529 )   Save

    BACKGROUND: In recent years, it has been a hot topic that stem cell transplantation is used to improve cardiac insufficiency after acute myocardial infarction by inducing regeneration of cardiomyocytes in the infarction regions.
    OBJECTIVE: To observe the effect of rosuvastatin combined with umbilical cord blood mesenchymal stem cells transplantation on rat cardiac function after acute myocardial infarction.
    METHODS: Forty-five Sprague-Dawley rats were enrolled to prepare myocardial infarction models by ligaturing the left anterior descending coronary artery. Then they were equivalently divided into model group, transplantation group and combination group. At 7 days after modeling, rats in the combination group were given injection of 300 μL umbilical cord blood mesenchymal stem cells (15.0×108) via the tail vein and by gavage once a day for 28 days with 1 mg/kg rosuvastatin; rats in the transplantation group and model group were injected with 300 μL umbilical cord blood mesenchymal stem cell suspension through the tail veins or the same amount of LG-DMEM medium, respectively, followed by intragastrical administration of the same amount normal saline. At 5 weeks after modeling, indexes of cardiac function, level of plasma Lp-PLA2 and heat shock protein 70 in the infarction regions were detected by color Doppler ultrasound, enzyme-linked immunosorbent assay and western blot assay, respectively. In addition, pathological changes of myocardial tissues were observed using hematoxylin-eosin staining.
    RESULTS AND CONCLUSION: Left ventricular ejection fraction and left ventricular end-systolic pressure were significantly higher in the combination group than in the transplantation group as well as higher in the transplantation group than the model group (P < 0.05); compared with the transplantation group, left ventricular end-diastolic pressure was significantly decreased in the combination group, but significantly increased in the model group (P < 0.05); the number of cardiomyocytes in the infarction regions was significantly higher in the combination group than the other groups. Additionally, expression of heat shock protein 70 in the infarction regions was significantly increased in the combination group (P < 0.05). To conclude, rosuvastatin combined with umbilical cord blood mesenchymal stem cell transplantation can significantly improve rat cardiac function after 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

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    Human umbilical blood cord mesenchymal stem cell transplantation combined with injection of ganglioside for cerebral palsy
    Yang Zi-jin, Guo Jia-li, Lu Si-guang, Gao Chang-long, Li Hong-mei, Feng Yue
    2016, 20 (19):  2803-2809.  doi: 10.3969/j.issn.2095-4344.2016.19.010
    Abstract ( 367 )   PDF (944KB) ( 512 )   Save

    BACKGROUND: In recent years, some studies have demonstrated that ganglioside can promote survival and differentiation of umbilical blood cord mesenchymal stem cells in vitro.
    OBJECTIVE: To observe the effect of injection of human umbilical blood cord mesenchymal stem cells and ganglioside into rat lateral ventricles on neurological functional recovery from cerebral palsy.
    METHODS: Totally 60 cerebral palsy neonatal rats were delivered from pregnant rats which were modes were given intraperitoneal injection of lipopolysaccharide for 2 successive days on day 17 of gestation. Then those neonatal rats were randomly divided into five groups, including model group (n=10), sham transplantation group (n=10), stem cell transplantation group (n=18), ganglioside group (n=10) and combination group (n=12). Under stereotaxic instrument, umbilical blood cord mesenchymal stem cells or ganglioside were injected into left lateral ventricles of the rat brain, respectively, and the sham transplantation group was given the same volume of phosphate buffered saline. Two rats from the stem cell transplantation group were put to death for immunofluorescence staining at 7, 14, 21 and 28 days after transplantation, respectively, and two rats in the combination group were killed for immunofluorescence staining at 14 days. Besides, all rats were underwent neurologic evaluation at 28 days after transplantation.
    RESULTS AND CONCLUSION: The umbilical blood cord mesenchymal stem cells could survive, migrate and differentiate, which mainly distributed in the lateral ventricle, hippocampus and cortex. At 14 days after transplantation, positive expressions of BrdU and glial fibrillary acidic protein in the combination group were significantly higher than those in the stem cell transplantation group (P < 0.05). In addition, compared with the model group, the holding time significantly prolonged and foot error times significantly decreased in the latter three groups (P < 0.05), as well as in the combination group compared with the stem cell transplantation and ganglioside groups (P < 0.05). These results indicate that umbilical blood cord mesenchymal stem cells and ganglioside can both improve neurological function of rats with cerebral palsy. Given that ganglioside can promote survival and differentiation of umbilical blood cord mesenchymal stem cells in vivo, the combined transplantation is preferred.

     

    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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    Propofol pretreatment combined with umbilical blood mesenchymal stem cell transplantation improves cerebral ischemia-reperfusion injuries
    Hou Shao-ke, Hao Li-na, Wei Jiao, Zhou Ya-jing
    2016, 20 (19):  2810-2816.  doi: 10.3969/j.issn.2095-4344.2016.19.011
    Abstract ( 341 )   PDF (1025KB) ( 401 )   Save

    BACKGROUND: As propofol has a neuroprotective effect, and umbilical cord blood mesenchymal stem cells have a high differentiation potential, their combination will have a better therapeutic effect on cerebral ischemia-reperfusion injury.
    OBJECTIVE: To study the effect of propofol pretreatment combined with umbilical blood mesenchymal stem cell transplantation on cerebral ischemia-reperfusion injury in rats.
    METHODS: Sixty-three Sprague-Dawley rats were randomized into model, propofol, and combined group (n=21 per group). Rat models of cerebral ischemia-reperfusion injury were made using ligation of the middle cerebral artery occlusion in the three groups. Rats in the combined group were given 100 mg/kg propofol injection at 1 day before injury and injection of umbilical blood mesenchymal stem cells via the tail vein (0.5 mL, 2×109/L).
    RESULTS AND CONCLUSION: Compared with the model group, the neurological function was improved significantly in the propofol and combined group, especially in the latter one, presenting with a remarkable mitigation in brain injury and an increased level of survivn mRNA in the rat hippocampus. The content of serum malondialdehyde was lower but the activity of superoxide dismutase was higher in the combined group compared with the propofol group. These findings indicate that propofol pretreatment combined with umbilical blood mesenchymal stem cell transplantation has better therapeutic effects than propofol pretreamtnet alone for improving cerebral ischemia-reperfusion injury 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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    Umbilical cord blood mesenchymal stem cell transplantation for acute intestinal ischemia-reperfusion injury
    Wang Bing-jie, Hu Yan-wei, Zhao Ye-fang, Zhang Shi-dong
    2016, 20 (19):  2817-2823.  doi: 10.3969/j.issn.2095-4344.2016.19.012
    Abstract ( 269 )   PDF (913KB) ( 390 )   Save

    BACKGROUND: Bone marrow mesenchymal stem cells can repair intestinal ischemia-reperfusion injuny by interfering inflammatory reactions after intestinal ischemia-reperfusion to protect intestinal barrier functions. In recent years, umbilical cord blood mesenchymal stem cells are gradually used as a substitute source of bone marrow mesenchymal stem cells.
    OBJECTIVE: To investigate the effects of umbilical cord blood mesenchymal stem cells on acute intestinal ischemia-reperfusion injury.
    METHODS: Umbilical cord blood mesenchymal stem cells were induced, isolated in vitro and tracked by CM-DiI fluorescent labeling. Sixty-three Sprague-Dawley rats were equivalently randomized into three groups: control group received normal saline enema, intestinal ischemia-reperfusion injury group with ethanol diluted trinitro-benzene-sulfonic acid and transplantation group administrated with 1×1010/L umbilical cord blood mesenchymal stem cell suspension via the tail vein at 1 hour after trinitro-benzene-sulfonic acid modeling. At 3 days after transplantation, colon tissues were removed in each group to observe pathological changes of the intestinal tract by hematoxylin-eosin staining. Besides, expression of leptin mRNA in the colon tissues and cyclooxygenase-2 in the mucosa were detected by RT-PCR and immunohistochemistry method, respectively.
    RESULTS AND CONCLUSION: Transplanted umbilical cord blood mesenchymal stem cells distributed in the intestinal lymphoid tissue and among glandular epithelial cells, suggesting that these stem cells might be involved in the process of intestinal ischemia-reperfusion injury repair. Compared with the control group, intestinal injury in the injury group was significantly aggravated, and most intestinal epithelial cells shed; and the transplantation group appeared to have significantly reduced intestinal damage and significantly less cell shedding. Expression of leptin mRNA was significantly higher in the injury group than the transplantation group followed by the control group, and there were significant differences among the three groups (P < 0.05). Additionally, expression of cyclooxygenase-2 in the injury group was significantly higher than that in the control group (P < 0.05); compared with the injury group, expression of cyclooxygenase-2 was significantly lower in the transplantation group (P < 0.05). To conclude, leptin and cyclooxygenase-2 may be involved in acute intestinal ischemia-reperfusion injury, and umbilical cord blood mesenchymal stem cell transplantation significantly lessens intestinal ischemia-reperfusion injury, which provides an experimental basis for human treating acute intestinal ischemic 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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    Gegen Qin Lian Tang combined with human amniotic mesenchymal stem cell transplantation for early diabetic retinopathy
    Liu Liang, Wang Jin-liang
    2016, 20 (19):  2824-2830.  doi: 10.3969/j.issn.2095-4344.2016.19.013
    Abstract ( 274 )   PDF (1041KB) ( 396 )   Save

    BACKGROUND: Human amniotic mesenchymal stem cells have the potential to differentiate into insulin-secreting cells and have immunomodulatory effects. Additionally, it has been clinically proved that Gegen Qin Lian Tang can lower blood sugar.
    OBJECTIVE: To investigate the effects of Gegen Qin Lian Tang combined with human amniotic mesenchymal stem cell transplantation on rat early diabetic retinopathy, by detecting pathological changes, levels of blood indicators and expressions of vascular endothelial growth factor in retinal tissues.
    METHODS: By intraperitoneal injection of streptozotocin, 42 Wistar rat models with diabetes mellitus were prepared and then randomly divided into model group, human amniotic mesenchymal stem cell transplantation group (cell transplantation group) and combination of human amniotic mesenchymal stem cells and Gegen Qin Lian Tang group (combination group). At 4 weeks after transplantation, levels of blood sugar and serum insulin in diabetic rats were detected; pathological changes of the retina in diabetic rats were observed by hematoxylin-eosin staining. Besides, expressions of CD45 and vascular endothelial growth factor mRNA were detected using immunohistochemistry staining and RT-PCR technology, respectively.
    RESULTS AND CONCLUSION: Compared with the model group, the levels of blood glucose and serum insulin in both cell transplantation and combination groups were significantly decreased and increased, respectively, and these changes were even more significant in the combination group (P < 0.05). In the model group, the retinal edema could be found, accompanied by structural disorder and irregular cell arrangement, while these retinal lesions were relatively milder in the cell transplantation group and significantly improved in the combination group. In addition, the CD45 expression and in the retina was highest in the model group subsequently followed by the cell transplantation group (P < 0.05) and lowest in the combination group (P < 0.05). Furthermore, compared with the model group, the expression of vascular endothelial growth factor mRNA was also significantly lower in the other two groups, especially in the combination group (P < 0.05). These data show that Gegen Qin Lian Tang combined with human amniotic mesenchymal stem cell transplantation improves the severity of diabetic retinopathy by inhibiting expressions of CD45 and vascular endothelial growth factor in the retinal tissues.

    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 mononuclear cell transplantation for cerebral hemorrhage
    Yang Yu-ye, Wang Jing-feng, Zhang Hong-yi, Wang Yue-wu, Yang Shu-quan
    2016, 20 (19):  2831-2837.  doi: 10.3969/j.issn.2095-4344.2016.19.014
    Abstract ( 372 )   PDF (1742KB) ( 444 )   Save

    BACKGROUND: It has been proved that bone marrow mononuclear cell transplantation can obviously improve neurological function of rats with cerebral hemorrhage.
    OBJECTIVE: To investigate the effects of transplanted bone marrow mononuclear cells on the neurological function and apoptosis in perihematomal brain tissues following cerebral hemorrhage in a rat model.
    METHODS: Twenty-four Sprague-Dawley rats were given stereotaxical injection of collagenase IV into the caudate nucleus to establish cerebral hemorrhage models in transplantation group (n=12) and model group (n=12), and then at 6 hours after cerebral hemorrhage, rats in these two groups were administrated 3x1010/L allograft bone marrow mononuclear cells and the same amount of PBS, respectively. Another 12 rats were given no interventions as control group. Neurological functions of rats were assessed at 1, 4, 8, 16 days after cerebral hemorrhage; pathological changes of the injury sites were observed at 16 days after transplantation; neuronal apoptosis rates in the perihematomal brain tissue were detected by flow cytometry at 2 and 4 days after transplantation.
    RESULTS AND CONCLUSION: The modified neurologic severity scores in the transplantation group were significantly lower than those in the model group at 8 and 16 days after cerebral hemorrhage (P < 0.05). In the control group, cells in each layer arranged closely with complete structure, and neurons and glial cells were in good shape; in the model group, perihematomal brain tissues were loose with intercellular gap, in which most neurons and glial cells became necrotic; in the transplantation group, cells in each layer arranged closely and regularly, and glial cell proliferation occurred. Besides, compared with the model group, the neuronal apoptosis rate in the transplantation group was significantly lower (P < 0.05). To conclude, bone marrow mononuclear cells can significantly enhance the neurological function recovery and reduce neuronal apoptosis in the brain of cerebral hemorrhage 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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    Neural stem cell transplantation for Alzheimer’s disease
    Zhang Ying, Yan Hong-juan, Guo Jian-hua, Luo Qiu-hua, Li Xiao-hui
    2016, 20 (19):  2838-2843.  doi: 10.3969/j.issn.2095-4344.2016.19.015
    Abstract ( 613 )   PDF (1777KB) ( 751 )   Save

    BACKGROUND: Neural stem cell transplantation has been used to treat a series of brain injury diseases, such as cerebral palsy, but its effect on Alzheimer’s disease is rarely reported.
    OBJECTIVE: To observe the effect of neural stem cell transplantation on the behavior and immune regulating system of Alzheimer’s disease rats. 
    METHODS: Thirty-five Sprague-Dawley rats were enrolled to make a postcerebral incision and given hippocampal injection of amanita phalloides acid to establish rat models of Alzheimer’s disease. Another 10 rats were only given hippocampal injection of normal saline after preparation of postcerebral skin incision as sham operation group. Then 32 successful rat models were randomly divided into two groups (n=16 per group): rats in experimental group were administrated hippocamal injection of 5×109/L allogeneic neural stem cell suspension; those in model group were given no injection. Five-day Morris water maze test was conducted at 4 weeks after transplantation. At 1 week after Morris water maze test, levels of interleukin-1 and interleukin-10 in the cerebral homogenate were detected, as well as pathological changes of brain tissues were observed in the three groups.
    RESULTS AND CONCLUSION: Compared with the model group, the abilities of cognition and memory were significantly higher in the sham operation group (P < 0.01), and the abilities of spatial learning and memory were significantly higher in the experimental group (P < 0.05, P < 0.01). Levels of interleukin-1 and interleukin-10 in the model group were significantly higher than those in the sham operation group (P < 0.01) but significantly lower than those in the experimental group (P < 0.01). Besides, the number of neurons in the model group was obviously less than that in the experimental and sham operation group. These results indicate that neural stem cell transplantation supplements and protects neurons against Alzheimer's disease in rats, thereby significantly 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

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    Variation of stromal cell-derived factor-1 level in a limbal stem cell deficiency model
    Zhang Wei, Huang Wen-zhi, Zhang Yu-ping
    2016, 20 (19):  2844-2849.  doi: 10.3969/j.issn.2095-4344.2016.19.016
    Abstract ( 384 )   PDF (1673KB) ( 346 )   Save

    BACKGROUND: How to promote homing of scarce limbal stem cells is the key to improving transplantation efficiency. And stromal cell-derived factor-1 plays an important role in the repair of retinal pigment epithelial cells injury.
    OBJECTIVE: To observe changes of stromal cell-derived factor-1 levels in rat and mouse limbal stem cell deficiency models in order to confirm whether these changes will induce peripheral blood stem cells homing into the cornea.
    METHODS: Eye models of limbal stem cell deficiency were prepared in rats and mice by cutting method, then peripheral blood samples were continuously drawn to detect the mass concentration of stromal cell-derived factor-1 by enzyme-linked immunosorbent assay within 1 month after modeling.
    RESULTS AND CONCLUSION: In the rat peripheral blood, the mass concentration of stromal cell-derived factor-1 started to increase at 24 hours after modeling and firstly peaked at the 3rd day; then it tended to decrease and reached a second peak lower than the first one at the 25th day; subsequently, it returned to the normal level. In the meanwhile, the mass concentration of stromal cell-derived factor-1 in the mouse peripheral blood, increased at 12 hours after modeling, peaked at the 3rd day, subsequently decreased until another peak lower than the former one presented at the 20th day, and finally it returned to the normal level. In conclusion, the level of stromal cell-derived factor-1 in the peripheral blood is significantly associated with limbal stem cells deficiency, which may play an important role in corneal injury repair.

     

    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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    Distribution and differentiation of embryonic liver stem cells in mice after intrahepatic transplantation
    Jiang Si-feng
    2016, 20 (19):  2850-2856.  doi: 10.3969/j.issn.2095-4344.2016.19.017
    Abstract ( 325 )   PDF (897KB) ( 385 )   Save

    BACKGROUND: As many factors can lead to liver injury, we attempt to use the “therapeutic liver regeneration” technology in clinical treatment of liver diseases by promoting liver regeneration.
    OBJECTIVE: To investigate distribution and differentiation of embryonic liver stem cells in mice after intrahepatic transplantation via a transplantation approach.
    METHODS: Liver injury models were prepared in 20 BALB/c mice, and then randomly equivalently assigned into two groups: 70% partial hepatectomy with intrahepatic transplantation with 1×105 embryonic liver stem cells in control group; therapeutic liver regeneration model plus intrahepatic transplantation with 1x105 embryonic liver stem cells in observation group. At 1 and 2 weeks after cell transplantation, the liver parenchyma of mice was observed. And at 2 weeks, both of the two groups underwent confocal immunofluorescence assay. Besides, blood samples of mouse tail vein were collected to detect levels of serum albumin.
    RESULTS AND CONCLUSION: At 1 week after cell transplantation, in the liver parenchyma, green fluorescence was sparsely distributed in the two groups, and the distribution density had no significant difference between the two groups; at 2 weeks after cell transplantation, hepatic cord-like structures appeared in the liver parenchyma of two groups, and the green fluorescence distribution in the control group was limited, but significantly expanded in the observation group. At 2 weeks after cell transplantation, positive albumin expression in the liver parenchyma was significantly higher in the observation group than in the control group, and there was no significant difference in levels of serum albumin between two groups (P > 0.05). To conclude, after transplantation of embryonic liver stem cells in the therapeutic liver regeneration model mice hepatocytes can be effectively integrated into the host hepatic plate, differentiate in the liver, and partially trigger the function of hepatocytes.

     

    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 cells on a novel composite scaffold: growth and differentiation
    Xing Ran, Chen Xu-yi, Zhu Xiang, Li Rui-xin, Tu Yue
    2016, 20 (19):  2857-2863.  doi: 10.3969/j.issn.2095-4344.2016.19.018
    Abstract ( 314 )   PDF (1097KB) ( 343 )   Save

    BACKGROUND: Neural stem cells with self-proliferation and differentiation potential are the ideal seed cells for central nervous tissue engineering. Although collagen and silk fibroin as biological scaffold materials have been widely used, both of them used alone have certain shortcomings. Is it possible to combine the two materials to build a novel neural tissue-engineered scaffold? What is the effect of this novel scaffold on the growth and differentiation of neural stem cells?
    OBJECTIVE: To observe the growth and differentiation of neural stem cells seeded onto the novel composite scaffold.
    METHODS: The rat embryonic neural stem cells were inoculated onto new composite scaffolds, and then, their growth and differentiation were observed by light microscopy and scanning electron microscopy. Neural stem cells were cultured in conventional suspension culture as control group. Cell counting kit-8 assay was used to detect viability of neural stem cells in the two groups. Three-dimensional composite scaffolds carrying neural stem cells were sliced into paraffin sections to observe the growth and differentiation of neural stem cells by hematoxylin-eosin staining and immunofluorescence staining.
    RESULTS AND CONCLUSION: Neural stem cells cultured on the new composite scaffold grew and differentiated well, and interconnected synapses were observed. Cell counting kit-8 assay showed that neural stem cells on the scaffold grew well, and the cell viability was significantly higher in the composite scaffold group than that in the control group (P < 0.05). Hematoxylin-eosin staining and immunofluorescence staining of paraffin sections further provided evidence for good growth and differentiation of neural stem cells on the scaffold. These results indicate that the novel composite scaffold with good biocompatibility benefits the growth and differentiation of neural stem cells, promising a favorable application prospect.

     

    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 mechanical factors on proliferation and differentiation of tissue- engineered stem cells
    Jiang Dong, Wang Ji-hong
    2016, 20 (19):  2864-2871.  doi: 10.3969/j.issn.2095-4344.2016.19.019
    Abstract ( 404 )   PDF (938KB) ( 438 )   Save

    BACKGROUND: With the development of tissue engineering technology and the deep research of tendon regeneration, many problems caused by traditional tendon transplantation will be solved.
    OBJECTIVE: To evaluate and prospect the effects of corresponding mechanical systems on seeding cells based on the characters of tendon, stem cells as well as mechanical systems.
    METHODS: CBM, CNKI, CqVip and PubMed databases were retrieved for reviews and articles related to tissue-engineered tendon published from January in 2005 to December in 2015. The keywords were “tissue engineering, tendon, tendon stem cell, mechanical stimulation and seeding cells” in Chinese and English, respectively. Finally a total of 63 articles were selected for overview.
    RESULTS AND CONCLUSION: As all kinds of stem cells and the corresponding mechanical systems exhibit advantages and disadvantages when applied for tissue engineering, it is advisable to make appropriate choices according to the research needs. Tendon stem cells show a broad application prospect in tendon regeneration, which will grow into required tendon tissues through appropriate, accurate and gentle mechanical stimulations, thereby providing another alternative to improve the tendon healing.

     

    中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

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    Dental pulp stem cells in regenerative medicine: application and development
    Ma Zi-yang, Guo Xiao-xia
    2016, 20 (19):  2872-2878.  doi: 10.3969/j.issn.2095-4344.2016.19.020
    Abstract ( 423 )   PDF (798KB) ( 1153 )   Save

    BACKGROUND: Dental pulp stem cells are characterized by multi-lineage differentiation and proliferation abilities and are easy to obtain, so they are becoming an issue of concern in regenerative medicine.
    OBJECTIVE: To provide clues and direction for further study by analyzing progress of domestic and overseas research on dental pulp stem cells, and summarizing their application in regenerative medicine.
    METHODS: The “dental pulp stem cell, regenerative medicine, tissue engineering” in Chinese and English served as the search terms to search articles related to dental pulp stem cells and regenerative medicine, published from 2000 to 2015 in Medline, PubMed, CNKI, Wanfang and Cqvip databases. Totally 46 articles were selected for overview. 
    RESULTS AND CONCLUSION: Dental pulp stem cells, which hold the capacity of self-renewal and multi-lineage differentiation, are relatively easy to obtain, and exhibit a great potential in regenerative medicine. The research of dental pulp stem cells in repairing bone defects has entered the clinical trial phase, but the research of cell differentiation into other tissues is still in basic trial phase and needs further development.

     

    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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    Development of mesenchymal stem cell homing
    Deng Rong-rong, Xie Yi-min, Xie Lin
    2016, 20 (19):  2879-2888.  doi: 10.3969/j.issn.2095-4344.2016.19.021
    Abstract ( 1121 )   PDF (1064KB) ( 1618 )   Save

    BACKGROUND: As a kind of adult stem cells with low immunogenicity, mesenchymal stem cells are able to differentiate into different cell lineages in the treatment of many diseases. Moreover, mesenchymal stem cells have been extensively used in many fields such as stem cell transplantation, immune therapy, and tissue engineering.
    OBJECTIVE: To review the progress of homing mechanism and the strategies to promote mesenchymal stem cell homing, thus providing a theoretical basis for transplanting mesenchymal stem cells safely and efficiently.
    METHODS: The CNKI and PubMed databases were retrieved by computer for articles regarding mesenchymal stem cell homing published from 2000 to 2016, including reviews, basic and clinical studies. The key words were “mesenchymal stem cells, homing” in Chinese and English, respectively. Then 74 papers were suitable for final analysis.
    RESULTS AND CONCLUSION: Mesenchymal stem cell homing needs further research, especially the molecular mechanism of cell mobilization. Therefore, basic research about mesenchymal stem cells should be further developed, and a standardized homing system should be established in vitro. In addition, it is of great significance to study the in vivo effects of transplanted gene-transfected 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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