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07 October 2012, Volume 16 Issue 41 Previous Issue    Next Issue

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Purification, cryopreservation and tumorigenicity of rat bone marrow mesenchymal stem cells Tong Ming-hua, Lei Xiang-li, Wang Ya-ping, Kong Xiang-ping, Luo Xian-rong 2012, 16 (41):  7601-7606.  doi: 10.3969/j.issn.2095-4344.2012.41.001 Abstract ( 238 )   PDF (602KB) ( 556 )   Save BACKGROUND: Bone marrow mesenchymal stem cells are restricted in clinical application due to low contents in bone marrow, easy to lose stem cell potential during long-term in vitro culture, and unclear safety after in vivo transplantation. OBJECTIVE: To explore an optimal isolation, purification and cytopreservation method of rat bone marrow mesenchymal stem cells and observe the tumorigenicity after implanting the in vitro cultured cells into nude mice. METHODS: Bone marrow mesenchymal stem cells were obtained from bone marrow cavity, purified and cutured by three different methods. The culture-expanded bone marrow mesenchymal stem cells were cryopreserved with cytoprotectant solution containing different serum concentrations at -80 ℃. Following recovery from cryopreservation, cell growth capacity and adiogenic capacity were determined. Passage 3 and 15 bone marrow mesenchymal stem cells were injected into the muscle and liver of nude mice to observe the neoplasia in local site. RESULTS AND CONCLUTION: The purity of bone marrow mesenchymal stem cells obtained by differential adhesion combined with 24-hour first exchange method was higher compared to other control groups, and there was no significant difference in cell proliferation capacity between groups. Bone marrow mesenchymal stem cells cryopreserved with 30% serum could ensure the viability, proliferation ability and stem cell characterisitics and multi-differentiation potential. Bone marrow mesenchymal stem cells might keep stem cell properties until to the 15th generation. After 45 days of transplantation to nude mice, no neoplasia was found in the local site. The muscle tissue had normal structure. Passage 15 bone marrow mesenchymal stem cells could survive and proliferate in the liver with no hetermorphism and invasion to the surrounding liver tissue. Related Articles | Metrics

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Establishment of rat bone marrow mesenchymal stem cell lines stably transfecting enhanced green fluorescent protein Li Yang, Tian Yu, Wu Zhi-chao, Sun Jia-xing, Wang Jian-jiao 2012, 16 (41):  7607-7611.  doi: 10.3969/j.issn.2095-4344.2012.41.002 Abstract ( 250 )   PDF (453KB) ( 480 )   Save BACKGROUND: The use of mesenchymal stem cells or therapeutic factor-containing stem cells is a promising therapeutic method to selectively kill tumor cells. OBJECTIVE: To establish rat bone marrow mesenchymal stem cell lines stably transfecting enhanced green fluorescent protein. METHODS: The lentiviral plasmid pVector-EGFP with the packaging plasmid pHelper 1 and the envelop plasmid pHelper 2 were co-transfected into 293T cells. The lentiviral titer was detected by real-time, fluorescence-based quantitative PCR. SD rat bone marrow mesenchymal stem cells during logarithmic phase were seeded in 24-well plates and exposed to lentiviral vector coding for EGFP reporter gene at multiplicity of infection (MOI) of 0, 5, 10, 15 and 20, respectively. EGFP expression and transfection efficiency were determined 72 hours later. RESULTS AND CONCLUSION: The lentiviral vector system carrying EGFP was successfully tranfected into 293T cells with a lentivirual titer of 1×108 TU/mL. At 2-3 days after rat bone marrow mesenchymal stem cells were transfected with packaged lentiviral vector coding for EGFP reporter gene, EGFP expression was detected in each well. MOI values ranged from 0 to 10. EGFP-expressing cells were gradually increased (P < 0.05). Over 70% cells were EGFP-positive at 3 days after transfection by lentiviral vector at MOI of 10. A significant dose-response was observed with increasing lentiviral titer for MOI 0 to 10 (P < 0.05), whereas the ratio of transfection decreased with increasing lentiviral liter for MOI 10 to 20. These findings suggest that lentivirus is an ideal vector for gene transduction and it can transduce an exogenous gene into rat bone marrow mesenchymal stem cells with over 70% efficency at the MOI of 10 to establish rat bone marrow mesenchymal stem cell lines stably transfecting EGFP. References | Related Articles | Metrics

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Effects of cadmium chloride on micronucleus rate and chromosome aberration rate in mouse bone marrow mesenchymal stem cells Liu Yong-qi, Da Rui, Dou Juan-juan, Su Ju-ping, Yan Chun-lu 2012, 16 (41):  7612-7616.  doi: 10.3969/j.issn.2095-4344.2012.41.003 Abstract ( 274 )   PDF (433KB) ( 457 )   Save BACKGROUND: There have been no reports describing the effects of cadmium chloride on heredity of bone marrow mesenchymal stem cells (BMSCs) OBJECTIVE: To investigate the effects of cadmium chloride on micronucleus rate and chromosome aberration rate in mouse BMSCs. METHODS: Mouse BMSCs were cultured by adherence method. Morphology of BMSCs was observed in the blank control group and cadmium chloride-induced group. The micronucleus rate and chromosome aberration rate in mouse BMSCs were determined. RESULTS AND CONCLUSION: Under the microscope, cells in the blank control group grew adherently like fibroblasts, with transparent soma and strong refraction; in the cadmium chloride-induced group, cells were retracted, somas became small and round, and were weakly adherent, and suspension cells increased. The micronucleus rate and chromosome aberration rate in mouse BMSCs were significantly increased in the cadmium chloride-induced group than in the blank control group (P < 0.05). These findings suggest that cadmium chloride can cause damage to BMSC genetic material. Related Articles | Metrics

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Effects of various volume fractions of umbilical cord serum on the proliferation ability of human bone marrow mesenchymal stem cells Lei Xiao-yu, Zhou Xin-Fu, Luo Zi-mian, Liu Li-hua, Liu Kang 2012, 16 (41):  7617-7620.  doi: 10.3969/j.issn.2095-4344.2012.41.004 Abstract ( 245 )   PDF (377KB) ( 389 )   Save BACKGROUND: Culture of bone marrow mesenchymal stem cells (BMSCs) after preparation of umbilical cord serum has become an increasing research area. However, there have been no reports regarding the effects of different volume fractions of umbilical cord serum on the proliferation ability of BMSCs. OBJECTIVE: To compare the effects of different volume fractions of umbilical cord serum on the proliferation ability of BMSCs. METHODS: Twenty bone marrow samples were obtained under aseptic conditions. The mononuclear cells were extracted by density gradient centrifugation method, and cultured with 5%, 7.5%, 10%, 15%, 20% umbilical cord serum + DMEM/F12 culture medium respectively. Passage 3 BMSCs were included in this study. MTT assay was used to investigate the effects of different volume fractions of umbilical cord serum on the proliferation ability of BMSCs. RESULTS AND CONCLUSION: Compared with the 15%, 20% volume fractions of umbilical cord serum + DMEM/F12 medium groups, the proliferation ability of BMSCs was significantly stronger than in the 5%, 7.5%, 10% volume fractions of umbilical cord serum + DMEM/F12 medium groups (P < 0.05). These findings suggest that the proliferation ability of BMSCs is more active at low volume fractions of human umbilical cord serum culture system. References | Related Articles | Metrics

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Isolation and preparation of Flk-1+ mesenchymal stem cells from mice and its biological characteristics Zhu Xi-shan, Jiang Ni, Song Yu-guang, Ren Jun 2012, 16 (41):  7621-7624.  doi: 10.3969/j.issn.2095-4344.2012.41.005 Abstract ( 252 )   PDF (416KB) ( 359 )   Save BACKGROUND: The bone marrow mesenchymal stem cells can be widely used, the three-lineage differentiation capacity and immune induction characteristics make it widely used in regenerative medicine. OBJECTIVE: To investigate the new method to isolate the Flk-1+ mesenchymal stem cells from mouse bone marrow, and to detect the biological characteristics of Flk-1+ mesenchymal stem cells. METHODS: The mononuclear cells were isolated from the bone marrow and the original mesenchymal stem cells were in vitro cultured and amplified. The cell cycle, phenotype and multilineage differentiation capacity were detected. RESULTS AND CONCLUSION: The original mesenchymal stem cells derived from mice bone marrow were fibrolbast like, and most of them stayed in G0/G1 phase. Cell phenotype test showed they could highly express Flk1, CD13, CD29, CD44; the bone marrow-derived Flk-1+ mesenchymal stem cells under light microscope showed a fibroblast-like or polygonal adherent growth, and under certain circumstances, they exhibited osteogenic, adipogenic and chondrogenic differentiation. Bone marrow-derived Flk-1+ mesenchymal stem cells have multilineage differentiation potential. Related Articles | Metrics

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Induced differentiation of human umbilical cord mesenchymal stem cells into chondrocytes in two kinds of culture systems Liu Jing, Xu Chao, Yu Li, Luo Er-mei, Zheng Yin-li, Wu Ying-hua, Chen Jian-ye, Zhou Zi-peng, Tang Ming-qiao 2012, 16 (41):  7625-7630.  doi: 10.3969/j.issn.2095-4344.2012.41.006 Abstract ( 227 )   PDF (555KB) ( 517 )   Save BACKGROUND: The methods of inducing mesenchymal stem cells to differentiate into chondrocytes under non-scaffold condition include high-density pellet culture, in vitro monolayer culture and co-culture with chondrocytes. High-density pellet culture and in vitro monolayer culture are widely concerned due to simple operation. OBJECTIVE: To find a better method of inducing human umbilical cord mesenchymal stem cells into chondrocytes in non-scaffold culture in vitro. METHODS: Human umbilical cord mesenchymal stem cells were isolated and cultured by tissue block method, and cell surface antigens were identified by flow cytometry. Human umbilical cord mesenchymal stem cells were cultured with chondrogenic media and stained with alcian blue. The production of matrix was estimated from the determination of hydroxyproline content using alcian blue method. Expressions of glycosaminoglycan, type Ⅱ collagen and Sox-9 were assayed by real-time fluorescence quantitative PCR. RESULTS AND CONCLUSION: The phenotype of cultured human umbilical cord mesenchymal stem cells was CD105+/CD29+/CD44+/ CD31-/CD34-/CD40-/CD45-/HLA-DR-. Human umbilical cord mesenchymal stem cells weakly expressed chondrocyte markers, which strongly expressed glycosaminoglycan and type Ⅱ collagen after chondgenic induction, and the expressions were higher in cells cultured in pellet culture system than in monolayer culture system. Real-time quantitative PCR results demonstrated that cells expressed higher level of glycosaminoglycan, type Ⅱ collagen and Sox-9 after induction than that before induction, and the expression levels were higher in cells cultured in pellet culture system than in monolayer culture system. The results revealed that human umbilical cord mesenchymal stem cells cultured in pellet culture system are more conducive to differentiate into chondrocytes than those cultured in monolayer culture system Related Articles | Metrics

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CM-Dil combined with DAPI label cellular membrane and nucleus of human umbilical cord blood mononuclear cells He Jin-ying, Yang Li-min, Ma Yu-zhen, Sun Wen-fang, Cen Yao, Yao Xing-yu 2012, 16 (41):  7631-7636.  doi: 10.3969/j.issn.2095-4344.2012.41.007 Abstract ( 273 )   PDF (647KB) ( 1040 )   Save BACKGROUND: Chlormethylbenzamido-1, 1-dioctadecyl-3, 3, 3’, 3’-tetramethylin-docarbocyamine (CM-DiI) and 4',6-diamidino-2-phenylindole (DAPI) are two kinds of tracers which are commonly used for labeling cellular membrane and nucleus respectively. OBJECTIVE: To investigate the feasibility of CM-DiI combined with DAPI labeling human umbilical cord blood mononuclear cells and to observe the changes in morphology and activities of the cells cultured in vitro after double labeling. METHODS: The freshly isolated mononuclear cells derived from human umbilical cord blood cells were double labeled with CM-DiI and DAPI, and the double labeled human umbilical cord blood cells were cultured in vitro, the changes of the morphology was observed under inverted phase contrast microscope, the activity of the cells at different times was detected though Trypan blue staining, at the same time, human umbilical cord blood cells double labeled by CM-DiI and DAPI were counted under the inverted fluorescence microscope at different times. RESULTS AND CONCLUSION: The membrane and nucleus of human umbilical cord blood cells present red and blue fluorescence respectively under different wavelengths with fluorescence microscope after double labeling by CM-DiI and DAPI for 15 minutes. The CM-DiI/DAPI double labeled human umbilical cord blood cells were in vitro cultured for 1, 3, 7, 14 and 21 days, and there was no significant difference in the number of positive cells double labeled by CM-DiI and DAPI at different time points under fluorescence microscope. The survival rates were 95.6%-98.8%. There was no significant difference in cell morphology between the in vitro cultured double labeled cells and the cells without labeling, and the growth state, adherent ability and cell proliferation ability of the double labeled cells were not changed. Human umbilical cord blood mononuclear cells can be effectively labeled by CM-DiI and DAPI at the same time. Both of the tracers have slower fluorescence decay and non-toxic adverse reactions on the living cells and are suitable for labeling the stem cells. Related Articles | Metrics

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Effect of platelet lysate on osteogenic differentiation of human umbilical cord derived mesenchymal stem cells in vitro Lü Jiang-tao, Tian Shao-qi, Sun Kang, Zhang Ji-hua, Zhang Ying-ying, Zhang Cai-long, Liu Shi-hai, Feng Xue-tao 2012, 16 (41):  7637-7641.  doi: 10.3969/j.issn.2095-4344.2012.41.008 Abstract ( 259 )   PDF (639KB) ( 484 )   Save BACKGROUND: Platelet lysate is the pyrolysis products of concentrated platelet which contains many active ingredients. Studies have shown that the active ingredients can promote the proliferation and differentiation of mesenchymal stem cells, but the effect of platelet lysate on the osteogenic differentiation of mesenchymal stem cells is not clear. OBJECTIVE: To study the intervention effect of platelet lysate in osteogenic differentiation of human umbilical cord derived mesenchymal stem cells in vitro. METHODS: Human umbilical cord derived mesenchymal stem cells isolated by collagenase digesting were cultured and amplified in vitro. Flow cytometer was used to detect the cell surface markers. The experiment was divided into four groups: mineralization medium group, platelet lysate group, mineralization medium-platelet lysate group and control group. The cells were cultured with corresponding induction medium for 2 weeks, then alizarin red staining was used to identify the oxteoblast, and alkaline phosphatase activity was used to compare the differentiation activity. RESULTS AND CONCLUSION: Flow cytometer results showed that CD34, CD45 and HLA-DR were negative in the human umbilical cord derived mesenchymal stem cells while CD44, CD105 and CD146 were positive. Alizarin red staining was positive in three induced groups with no obvious differences, and it was negative in the control group. The alkaline phosphatase activity level in three induced groups was significantly higher than that in the control group (P < 0.05), and the alkaline phosphatase activity level in mineralization medium-platelet lysate group was significantly higher than that in the mineralization medium group and platelet lysate group (P < 0.05). Results show that 5% platelet lysate alone can induce human umbilical cord derived mesenchymal stem cells to differentiate into osteoblasts in vitro. The induction will be more efficient when combining 5% platelet lysate with mineralization medium. Related Articles | Metrics

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Human embryonic stem cells seeded in rat tail collagen-coated culture plates directly differentiate into vascular endothelial cells Luo Min, Geng Ju-min, Liang Dao-ming, Hu Zhi-xing 2012, 16 (41):  7642-7646.  doi: 10.3969/j.issn.2095-4344.2012.41.009 Abstract ( 274 )   PDF (457KB) ( 583 )   Save BACKGROUND: Human embryonic stem cells (hESCs) can be induced to differentiate into vascular endothelial cells in vitro, which be involved in the formation of the blood vessels. Therefore, hESCs is one of the promised cell resources for vascular tissue engineering. OBJECTIVE: To establish a method to directly induce the hESCs to differentiate into vascular endothelial cells. METHODS: hESCs H1 was cultured on mouse embryonic fibroblast (MEF) in serum-free ESC medium. After passaged, H1 cell clumps were transferred to rat tail collagen-coated culture plates, and then cultured on EGM-2 endothelial medium added with endothelial cell growth supplements and growth factors after adherent cultured for 24-48 hours. RESULTS AND CONCLUSION: Cells cultured on EGM-2 endothelial medium gradually expressed endothelial-specific genes VEGFR-2, PECAM1, vWF, CD34, VE-cadherin and GATA-2. The differentiated cells could express endothelial- specific genes VE-cadherin and CD31. The differentiated cells could uptake low density lipoprotein. hESCs on tail collagen-coated culture plates could directly differentiate into vascular endothelial cells in EGM-2 endothelial medium. The establishment of endothelial differentiation system will lay a foundation for further researches on the effects of extracellular matrix on inducing hESCs to differentiate vascular endothelial cells and its molecular mechanisms. Related Articles | Metrics

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Temporal specificity of differentiation of rat embryonic neural stem cells cultured in vitro Huang Fei, Zhao Ji-tong, Shen Qiang 2012, 16 (41):  7647-7650.  doi: 10.3969/j.issn.2095-4344.2012.41.010 Abstract ( 248 )   PDF (460KB) ( 439 )   Save BACKGROUND: There have been few studies describing the characteristics of self differentiation of neural stem cells cultured in vitro. OBJECTIVE: To investigate the temporal specificity of differentiation of rat embryonic neural stem cells cultured in vitro. METHODS: Neural stem cells were isolated from rat embryonic cerebral cortex and cultured in serum-free medium. Neural stem cells cultured for different time were induced to differentiate and the change of differentiation properties was detected. RESULTS AND CONCLUSION: Neural stem cells cultured in vitro were induced to differentiate predominately into neurons at early phase. As the increase of incubation time, the proportion of neural stem cells differentiating into neurons was decreased, and the proportion of neural stem cells differentiating into astrocytes was increased. However, there was no obvious correlation between the differentiation potential of neural stem cells into oligodendrocytes and incubation time. These findings suggest that as the increase of incubation time, the differentiation potential of neural stem cells into neurons was reduced and the differentiation potential of neural stem cells into astrocytes was enhanced; there was no obvious correlation between the differentiation potential of neural stem cells into oligodendrocytes and incubation time. Related Articles | Metrics

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Effect of adipose-derived stem cells modified with hypoxia-inducible factor-1 alpha gene on acute renal injury induced by cisplatin in mice Wang Wei-wei, Jiang Yan, Wang Wei, Cheng Jin, Zhang Jin-yuan 2012, 16 (41):  7651-7657.  doi: 10.3969/j.issn.2095-4344.2012.41.011 Abstract ( 233 )   PDF (709KB) ( 504 )   Save BACKGROUND: Stem cells transplantation provides a new way for the treatment of renal injury. The effect could be enhanced by transfecting target gene before transplantation. OBJECTIVE: To explore the effect of adipose-derived mesenchymal stem cells transfected with hypoxia-inducible factor-1α gene on the structure and function of the kidney in mice with acute renal injury induced by cisplatin. METHODS: The cisplatin-induced acute renal injury mice model was established by intraperitoneal injecting with 10 mg/kg cisplatin for 2 days. At 24 hours after modeling, adipose-derived mesenchymal stem cells suspension containing 1×105 adipose-derived mesenchymal stem cells or hypoxia-inducible factor-1α was administrated through tail vein, after 3 days, the blood and kidney samples were detected. The acute renal injury mice injected with 200 μL normal saline were regarded as model controls, and the normal mice were as normal controls. RESULTS AND CONCLUSION: After adipose-derived mesenchymal stem cells intervention, the serum creatinine and blood urea nitrogen level were decreased, and the pathological changes of renal tissues and apoptosis lesions of renal tubular epithelial cells were decreased, as well as the expression of inflammatory cytokines RANTES of kidney tissue and tumor necrosis factor-α, while the expression of interleukin-10 was increased; the hypoxia-inducible factor-1α modified adipose-derived mesenchymal stem cells had a positive effect on renal cell apoptosis and expression of inflammatory cytokines. The immunofluorescence staining showed that a small quantity of green fluorescent protein from adipose-derived mesenchymal stem cells was present in renal tissue, but transformatation to the renal tubular epithelial cells was not observed. The results show that adipose-derived mesenchymal stem cells transplantation can improve the damage of the kidney structure and function in acute renal injury, and the adipose-derived mesenchymal stem cells transfected with hypoxia-inducible factor-1α are more effective in repairing acute renal injury. Related Articles | Metrics

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Construction of tissue engineering trachea by in vitro and in vivo culture of adipose-derived stem cells in three-dimensional scaffolds Guo Xiang, Guan Xin, Zhao Heng 2012, 16 (41):  7658-7663.  doi: 10.3969/j.issn.2095-4344.2012.41.012 Abstract ( 231 )   PDF (681KB) ( 404 )   Save BACKGROUND: Trachea substitutes include autologous tissue flap, allogeneic tracheal transplant, artificial scaffolds and non-dynamic tissue transplant. However, they are hardly used in the clinic because of severe complications and difficult harvest. OBJECTIVE: To establish the tissue-engineered tracheal scaffold model by adipose-derived stem cells, poly-D,L-lactio-co-glycolic acid and poly(trimethylenecarbonate). METHODS: Adipose-derived stem cells of Sprague Dawley rats were isolated and cultured by tissue block method. Cell surface markers were evaluated by flow cytometry and multilineage differentiation, and the cells were implanted on the poly-D,L-lactio-co-glycolic-poly(trimethylenecarbonate) scaffold. After in vitro and in vivo culture, the adipose-derived stem cells were observed by immunohistochemistry and scanning electron microscope. RESULT AND CONCLUSION: Adipose-derived stem cells were spherical and well attached to the scaffolds with lamellipodia and conglobate connection after inoculated on the poly-D,L-lactio-co-glycolic-poly(trimethylenecarbonate) scaffold. After in vitro culture, the newborn capillary could be seen. The poly-D,L-lactio-co-glycolic- poly(trimethylenecarbonate) scaffold scaffolds with porous structure were highly biocompatible and non-cytotoxic as determined by adipose-derived stem cells adhesion and proliferation. Neotrachea fabricated with combined in vitro and in vivo culture system using poly-D,L-lactio-co-glycolic-poly(trimethylenecarbonate) scaffold and adipose-derived stem cells can meet the requirement of tracheal replacement, this model serve as a new way to engineer a vascularized trachea. Related Articles | Metrics

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Adipogenic differentiation of bone marrow mesenchymal stem cells from ovariectomized rats Liang Shi-zhen, Zhu Guo-xiong, Jin Yan, Wu Gao-yi, Sun Xi-qian 2012, 16 (41):  7664-7668.  doi: 10.3969/j.issn.2095-4344.2012.41.013 Abstract ( 271 )   PDF (528KB) ( 511 )   Save BACKGROUND: Under appropriate conditions, enhanced adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) can promote the formation of adipocytes in vivo. OBJECTIVE: To investigate the influence of ovariectomy on rat body mass, and to analyze the pathological mechanism from the point of functional changes of BMSCs. METHODS: Healthy female Sprague-Dawley rats of similar ages, body mass and from the same batch were selected and randomly divided into two groups. In the ovariectomy group, osteoporosis was induced by resection of bilateral ovaries. In the sham-surgery group, partial adipose tissue adjacent to bilateral ovaries was resected. Three months later, bone quality of the femur and body mass were determined by Micro-CT examination in each group. Rat BMSCs were isolated and cultured in vitro till passage 5. After adipogenic inducation, oil red O staining of lipid drops was used to detect the adipogenic ability of BMSCs; reverse transcription-polymerase chain reaction was used to detect the mRNA expression of peroxisome proliferator-activated receptor gamma and lipoprotein lipase, BMSCs adipogenesis related proteins. RESULTS AND CONCLUSION: Rat osteoporosis model was successfully constructed 3 months after ovariectomy. Compared to the sham-surgery group, the femoral bone quality was significantly decreased, body mass was significantly increased, and adipogenic differentiation of BMSCs was significantly enhanced in the ovariectomy group (P < 0.05). These findings suggest that enhanced adipogenic differentiation of BMSCs is likely to be the important cause for increased adipogenesis in vivo and subsequent obesity-like change of body mass in rats after ovariectomy. Related Articles | Metrics

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Chemokine expression in the ovarian tumor stem cells and cell invasion and metastasis Sun Hui 2012, 16 (41):  7669-7672.  doi: 10.3969/j.issn.2095-4344.2012.41.014 Abstract ( 274 )   PDF (352KB) ( 421 )   Save BACKGROUND: The method that using chemokine or chemotactic polypeptide to recruit immune effector cells to converge in tumor cells to arise anti-tumor immunity is expected to become a new strategy for tumor treatment. OBJECTIVE: To discuss the effects of chemokine expression on the invasion and metastasis in the ovarian tumor stem cells for giving new ideas of ovarian cancer research. METHODS: Expression of CXCR4 in ovarian cancer cell line SW626 was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Transwell room was used to detect the effects of CXCL12 and CXCL12/CXCR4 on chemotaxic invasion and metastasis of SW626 cells. RESULTS AND CONCLUSION: RT-PCR confirmed that there was CXCR4 expression in the SW626 cell lines, but no expression was found in the Anglne cells. CXCR4 expression could increase the number of ovarian tumor stem cells subjected to chemotactic invasion and metastasis, while CXCR4 addition could inhibit above-mentioned effects. CXCR12, a chemotatic factor of ovarian tumor stem cells, can promote the chemotaxic activity and invasion and metastasis activity mediated by CXCR4 receptor. Related Articles | Metrics

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Induction of transplantation tolerance to fully mismatched skin allografts in mice by syngeneic hematopoietic stem cells Xu San-rong, Zhou Qing, Han Bo, Liu Xia, Wu Wei-jiang, Xu Hui-ling 2012, 16 (41):  7673-7676.  doi: 10.3969/j.issn.2095-4344.2012.41.015 Abstract ( 292 )   PDF (421KB) ( 390 )   Save BACKGROUND: The optimal effect of organ transplantation tolerance is to induce the specific immunologic tolerance to transplantation antigens. OBJECTIVE: To study the possibility of using syngeneic hematopoietic stem cells to reconstitute immune system and induce transplantation tolerance to fully mismatched skin allografts in mice. METHODS: BALB/c mouse marrow was taken. Syngeneic skin transplantation was performed taking C57BL/6 mice as donors and BALB/c mice as recipients. Thirty-two BALB/c mice were randomly divided into four groups: control, cyclosporin A, cyclosporin A + total body irradiation, cyclosporin A + total body irradiation, cyclosporin A + total body irradiation + bone marrow transplantation. RESULTS AND CONCLUSION: In the cyclosporin A + total body irradiation group, all mice died within 10 days and the counts of peripheral blood white blood cells were persistently declined. In the cyclosporin A + total body irradiation + bone marrow transplantation group, mice survived during the experimentation, the counts of peripheral blood white blood cells decreased to the bottom after 6 days of irradiation and then rebounded, and increased to the level of the cyclosporine A group after 21 days of irradiation (P > 0.05). The average survival time of skin grafts in the cyclosporin A + total body irradiation + bone marrow transplantation group was significantly longer than that in the other groups (P < 0.01). There were less infiltrated cells and less destructed tissue, and decreased proliferation of recipient mouse spleen cells in the cyclosporin A + total body irradiation + bone marrow transplantation group than in the other groups. These findings suggest that immune reconstitution with syngeneic bone marrow cells can obviously prolong the survival time of mouse skin allografts and induce donor-specific tolerance. Related Articles | Metrics

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Different transplantation approaches of bone marrow mesenchymal stem cells in a rat model of cirrhosis Wu Yu-zhuo 2012, 16 (41):  7677-7680.  doi: 10.3969/j.issn.2095-4344.2012.41.016 Abstract ( 261 )   PDF (442KB) ( 409 )   Save BACKGROUND: Transplantation of bone marrow mesenchymal stem cells can alleviate cirrhosis and improve liver function. OBJECTIVE: To investigate the differences among the curative effects of three transplantation approaches of mesenchymal stem cells (MSCs) in a rat model of cirrhosis. METHODS: 60 SD rats were randomly divided into five groups: normal, control, portal vein transplantation, hepatic artery transplantation and tail vein transplantation groups. The rat model of CC14 and alcohol-induced cirrhosis was prepared in the latter four groups. Rats in the portal vein transplantation, hepatic artery transplantation and tail vein transplantation groups received 1×106 allogenic rat bone marrow mesenchymal stem cells via the portal vein, hepatic artery and tail vein, respectively. The control group rats received no transplantation. RESULTS AND CONCLUSION: Compared with the control group, the liver function of cirrhosis rats in the three transplantation groups was improved significantly, the levels of serum albumin and cholinesterase were significantly increased (P < 0.05), the levels of transaminases, bilirubin, prothrombin time, type IV collagen were significantly decreased, and the degree of cirrhosis was significantly alleviated (P < 0.05) at 4 weeks after transplantation in the three transplantation groups. The outcomes were significantly superior in the portal vein transplantation and hepatic artery transplantation groups than in the tail vein transplantation group (P > 0.05). These findings suggest that portal vein transplantation and hepatic artery transplantation can better alleviate the degree of cirrhosis and improve hepatic function than tail vein transplantation, and there is no significant difference between the former two groups. Related Articles | Metrics

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Bone marrow stem cell mobilization with stem cell factor and granulocyte colony-stimulating factor for treatment of renal ischemia/reperfusion injury Bi Ling-yun, Guo Jin-gang, Zhang Rui-xia, Zhao Jing-li, Liang Bin, Zhao De-an, Yang Da-sheng 2012, 16 (41):  7681-7687.  doi: 10.3969/j.issn.2095-4344.2012.41.017 Abstract ( 451 )   PDF (736KB) ( 530 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) can differentiate into the native cells of renal tissue to repair the injured renal tissue. Under normal circumstance, peripheral blood stem cells are limited in number, and mobilization of BMSCs can increase the number of peripheral blood stem cells. OBJECTIVE: To observe the therapeutic effects of BMSC mobilization on repair of ischemia/reperfusion-induced renal injury and on hypoxia inducible factor-1a and to investigate the mechanism by which BMSC mobilization repairs renal injury. METHODS: Sprague-Dawley rats were randomly allocated into four groups: In the control group, there was no treatment. In the model group, renal ischemia/reperfusion model was prepared. In the treatment group, 200 μg/kg per day stem cells and 50 μg/kg per day granulocyte colony-stimulating factors were subcutaneously administered in rat models of ischemia/reperfusion renal injury to mobilize BMSCs. In the treatment control group, normal rats received the same administration as rats in the treatment group. Drug administration was performed for a total of 5 successive days. At 5, 10, 17, 24, 31 days post-surgery, renal tissue was resected for pathological observation, and the expression level of CD34+ cells, hypoxia inducible factor-1a, vascular endothelial growth factor and heme oxygenase 1 was detected. RESULTS AND CONCLUSION: Application of stem cell factors combined with granulocyte colony-stimulating factors could significantly increase BMSCs in the injured renal tissue and alleviate the injury degree of renal tissue. BMSCs can increase hypoxia inducible factor-1a expression, vascular endothelial growth factor and heme oxygenase 1, which may be one of possible mechanisms by which BMSCs promote the repair of acute renal injury. BMSC mobilization can promote the expression of hypoxia inducible factor-1a system. Related Articles | Metrics

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Effect of stromal cell derived factor-1 on migration of bone marrow mesenchymal stem cells and bone healing Chen Wei, Chen Jian-mei, Zhang Xian-yu, Yao Xiao-dong, Xu Hao 2012, 16 (41):  7688-7693.  doi: 10.3969/j.issn.2095-4344.2012.41.018 Abstract ( 315 )   PDF (573KB) ( 394 )   Save BACKGROUND: The number and function of bone marrow mesenchymal stem cells in the fracture end are closely correlated with fracture healing. Stromal cell derived factor-1 can promote the chemotactic functions of bone marrow msenchymal stem cells. OBJECTIVE: To establish the left tibia fracture model by using the chimeric mice with stably expressing green fluorescence protein positive bone marrow mesenchymal stem cells, in order to evaluate the protective effect of stromal cell derived factor-1 on the migration of bone marrow mesenchymal stem cells and bone healing, and to explore the mechanism. METHODS: Green fluorescence protein positive bone marrow mesenchymal stem cells were harvested from bone marrow of green fluorescence protein transgenic C57BL mice by density gradient centrifugation method. The green fluorescence protein transgenic bone marrow mesenchymal stem cells combined with the non-adherent bone marrow cells in the male mice were transplanted after X-ray irradiation for establishing the chimeric mice model. After chimeric mice were established, left tibia fracture was made and induced with stromal cell derived factor-1 and stromal cell derived factor-1 antibody, and the control group was set. RESULTS AND CONCLUSION: The amount of bone marrow mesenchymal stem cells in stromal cell derived factor-1 group was higher than that in the control group and anti-stromal cell derived factor-1 group at 1, 3, 7 and 14 days after modeling (P < 0.05); the sum of callus in stromal cell derived factor-1 group were greater than that in the control group and anti-stromal cell derived factor-1 group at 14 and 21 days after modeling (P < 0.05); the average peak deformation of the fracture-healing site in the stromal cell derived factor-1 group at 28 days after modeling was greater than that in the control group and anti-stromal cell derived factor-1 group (P < 0.05); all the indicators above in the control group were greater than those in the anti-stromal cell derived factor-1 group. At 28 days after modeling in the stromal cell derived factor-1 group, the sum of callus was decreased (P < 0.05), the trabecular was integrated into the film and part of the marrow cavity was recanalized. The medullary cavity in the control group and anti-stromal cell derived factor-1 group was not recanalized. Stromal cell derived factor-1 can promote bone marrow mesenchymal stem cells to migrate into the fracture site and can accelerate fracture healing. Related Articles | Metrics

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Brain-derived neurotrophic factor promotes the directional differentiation of hypoglycemia neonatal rat hippocampal neural stem cells Tang Yuan-yuan, Li Zhen-yu, Yin Yan-yan, Ji Li-li, Wang Zhen-yu 2012, 16 (41):  7694-7697.  doi: 10.3969/j.issn.2095-4344.2012.41.019 Abstract ( 317 )   PDF (398KB) ( 415 )   Save BACKGROUND: There is evidence that brain-derived neurotrophic factor can maintain neuronal survival, influence neuronal migration, and promote the survival and differentiation of neural stem cells in vitro. OBJECTIVE: To investigate the effects of brain-derived neurotrophic factor on the directional differentiation of hypoglycemia neonatal rat hippocampal neural stem cells. METHODS: Neonatal 1-day hypoglycemia rat hippocampal tissue was harvested and subjected to primary culture, subculture and cell monoclonal culture. Then cells were divided into two parts: one part was used for identification of neural stem cells, and the other part was divided into three groups as per different concentrations of brain-derived neurotrophic factors (0, 100, 200 μg/L). Passage 4 cells were used for induced differentiation. Neuron-specific enolase immunofluorescence staining was used for neuronal identification and positive cells were counted. RESULTS AND CONCLUSION: After monoclonal culture, three groups of cells were positive for nestin. After induced differentiation, cells were positive for neuron-specific enolase and glial fibrillary acidic protein staining. Neural stem cells in the 100 μg/L and 200 μg/L groups grew faster, and produced higher proportion of neuron-specific enolase-positive cells than those in the 0 μg/L group (P < 0.05). But there was no significant difference between 100 μg/L group and 200 μg/L group. These findings suggest that brain-derived neurotrophic factor promotes the directional differentiation of hypoglycemia neonatal rat hippocampal neural stem cells. Related Articles | Metrics

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Transfection and expression of swine bone marrow mesenchymal stem cells hHCN2 gene cultured in vitro Min Rui, Li Jian-mei, Tao Si-ming, Zhang Xin-jin, Li Xian-hua 2012, 16 (41):  7698-7703.  doi: 10.3969/j.issn.2095-4344.2012.41.020 Abstract ( 219 )   PDF (561KB) ( 383 )   Save BACKGROUND: Recently, the pacemaker channel that modulates cardiac automaticity via the hyperpolarization activated caution current is the focus of this area. OBJECTIVE: To transfer the gene constructs CMV-hHCN2-3xHA-IRES-EGFP plasmid into the swine bone marrow mesenchymal stem cells (BMSCs) and to observe the expression of nucleic acid and protein in BMSCs. METHODS: BMSCs were separated, purified and proliferated in vitro by direct adherent or density gradient centrifugation combined with direct adherent method. The target gene CMV-hHCN2-3xHA-IRES-EGFP was transfected into BMSCs using lipofectamine 2000. RESULTS AND CONCLUSION: The growth speed of BMSCs cultured by using density gradient centrifugation was faster than that by direct adherent method. There was fluorescence of BMSCs under fluorescence microscope after the third generation of BMSCs transfected for 48 hours, there was no fluorescence in the control group. The transfection rate of the cells was 15%-20%. The expression of human hyperpolarization-activated cyclic nucleotide gated channel 2 (hHCN2) was observed on BMSCs by RT-PCR and Western blot. The growth speed of BMSCs cultured using density gradient centrifugation combined with direct adherent was faster than that by direct adherent method. The protein overexpression of hHCN2 gene was detected. Related Articles | Metrics

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Construction of human telomerase catalytic subunit gene lentiviral expression vector and preliminary study on transfection of human bone marrow stromal cells Chen Zhen-zhou 2012, 16 (41):  7704-7708.  doi: 10.3969/j.issn.2095-4344.2012.41.021 Abstract ( 352 )   PDF (476KB) ( 497 )   Save BACKGROUND: Bone marrow stromal cells (BMSCs) lack telomerase activity. Therefore, cell telomere length gradually shortens while proliferation in vitro, resulting in cellular senescence. This is an important factor that limits cell therapy application of BMSCs. OBJECTIVE: To construct human telomerase catalytic subunit (hTERT) gene lentiviral expression vector, and to investigate the feasibility of transfection of human BMSCs with lentivirus-mediated hTERT gene. METHODS: hTERT cDNA was obtained by PCR amplification from pReceiver-M02-hTERT. The coding sequence of hTERT was transferred into pDONR221 using BP recombination system, and then was transferred from pDONR221 into pLenti6.3/V5-DEST using LR clonase. The recombinant plasmid vector and packaging mixture were transfected into 293FT using Lipofectamine 2000 reagent, and the lentiviral particles were collected. ELISA was employed to detect the lentiviral titers. Q-PCR, Western blot and fluorescence immunocytochemistry were carried out to detect gene expression in human BMSCs after lentiviral transfecton. RESULTS AND CONCLUSION: The hTERT gene lentiviral expression vector was constructed successfully. The average virus physical titer is 1.07×1012 LP/L. Q-PCR confirmed that the hTERT gene expression level was significantly improved in modified human BMSCs. Western blot and immunocytochemistry confirmed the correct expression of hTERT protein in cells. The results suggest that transfection of human BMSCs with lentivirus-mediated hTERT gene can enhance hTERT expression. Related Articles | Metrics

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Effect of bone marrow mesenchymal stem cells on vascular endothelial growth factor expression in the aging heart of rats Li Yan-ju, Liu Yang, Guo Kun-yuan 2012, 16 (41):  7709-7712.  doi: 10.3969/j.issn.2095-4344.2012.41.022 Abstract ( 279 )   PDF (401KB) ( 375 )   Save BACKGROUND: Vascular endothelial growth factors secreted by bone marrow mesenchymal stem cells may exhibit therapeutic effects on the aging heart. However, whether the therapeutic effects are associated with bone marrow mesenchymal stem cells homing to the heart? OBJECTIVE: To investigate the effects of bone marrow mesenchymal stem cells on aging heart. METHODS: Rat bone marrow mesenchymal stem cells were harvested. Forty rats were randomly divided into four groups: normal, model, treatment and fluorescence labeling. Rats in the latter three groups were subcutaneously injected with D-galactose to establish aging heart rat models. Rats in the latter two groups were injected with non-labeled or carboxyflu-orescein diacetate succinimidyl ester-labeled bone marrow mesenchymal stem cells. Cell location, cardiac tissue factor expression were investigated. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells could arrive at the aging heart. Compared with the model group, the activity of superoxide dismutase in the serum and heart tissue was significantly increased, the level of malondialdehyde was significantly decreased (P < 0.05), and vascular endothelial growth factor expression in the cardiac tissue was significantly increased after treatment with bone marrow mesenchymal stem cells. These findings suggest that bone marrow mesenchymal stem cells can secrete vascular endothelial growth factors, which may be one of mechanisms by which bone marrow mesenchymal stem cells contribute to improvement of aging heart. Related Articles | Metrics

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Ginsenoside Rb1 reduced beta-amyloid peptide 25-35-induced hyperphosphorylation of Tau protein during the differentiation of neural stem cells Zhao Qing-xia, Li Bing-hua, Han Xue-fei, Xu Yan, Wu Guo-hua 2012, 16 (41):  7713-7716.  doi: 10.3969/j.issn.2095-4344.2012.41.023 Abstract ( 346 )   PDF (406KB) ( 836 )   Save BACKGROUND: Previous studies have demonstrated that ginsenoside Rb and beta-amyloid peptide 25-35 (Aβ25-35) can regulate the phospholation of Tan protein during the differentiation of neural stem cells (NSCs). Protein phosphatase 2A (PP2A) is closely related to hyperphosphorylation of Tau protein. OBJECTIVE: To investigate the effects of Aβ25-35 and ginsenoside Rb1 on phosphorylation level of Tau protein and activity of PP2A during the differentiation of NSCs. METHODS: NSCs were isolated from newborn rat hippocampus. After culture for 1 week, passage 3 NSCs were divided into three groups. In the control group, NSCs were further cultured for 36 hours without any medium added. In the Aβ group, after 24 hours of culture, Aβ25-35 was added for another 12 hours of culture. In the ginsenoside Rb1 froup, ginsenoside Rb1 was added for 24 hour pretreatment, and Aβ25-35 was added for another 12 hours of culture. The expression of Tau[pS396] and Tau[pS262] was tested by the immunofluorescent cytochemical staining and western blot method, and PP2A activity was tested by ELISA. RESULTS AND CONCLUSION: Cellular expression of Tau[pS396] and Tau[pS262] was detected during the differentiation of NSCs. In the Aβ group, cellular expression of Tau[pS396] and Tau[pS262] was up-regulated, and PP2A activity was not altered obviously. In the ginsenoside Rb1 group, cellular expression of Tau[pS396] and Tau[pS262] was down-regulated and PP2A activity was significantly increased. These findings suggest that during normal differentiation of NSCs, Tau protein was phosphorylated at a certain level, and ginsenoside Rb1 can alleviate Aβ25-35-induced hyperphosphorylation of Tau protein during the differentiation of NSCs by increasing PP2A activity. Related Articles | Metrics

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Tougu Xiaotong Granule water/alcohol extract-containing serum interferes the activity of bone marrow stromal stem cells Liu Bo-ling, Li Xi-hai, Liang Gui-qing, Chen Qi-yong, Wu Guang-wen, Li Zhao-hui 2012, 16 (41):  7717-7722.  doi: 10.3969/j.issn.2095-4344.2012.41.024 Abstract ( 248 )   PDF (514KB) ( 362 )   Save BACKGROUND: Bone marrow stromal stem cells have multilineage differentiation capacity and can differentiate into transparent chondrocytes, which can provide new thoughts for treatment of osteoarthritis. OBJECTIVE: To investigate the effects of Tougu Xiaotong Granule water/alcohol extract-containing serum on the activities of bone marrow stromal stem cells. METHODS: Tougu Xiaotong Granule water/alcohol extracts were prepared. A total of 108 SD rats were randomly divided into control group, Tougu Xiaotong Granule water extract group and Tougu Xiaotong Granule alcohol extract group. Then each group was divided into three subgroups as per low, medium, and high dose. Abdominal aorta blood was taken to prepare drug-containing serum. Passage 3 bone marrow stromal stem cells from 4-week-old SD rats were interfered with Tougu Xiaotong Granule water/alcohol extract-containing serum, and then cell activity was detected by MTT. Cell cycle was determined by flow cytometry. Cyclin D1 mRNA expression was detected by real-time PCR. Cyclin D1 protein expression was determined by western blot analysis. RESULTS AND CONCLUSION: After 48 hours of intervention by Tougu Xiaotong Granule water/alcohol extract, the ratio of cells in the G0/G1 period in the Tougu Xiaotong Granule water extract group and Tougu Xiaotong Granule alcohol extract group was significantly lower than that in the control group (P < 0.01), the ratio of cells in the G0/G1 period in the Tougu Xiaotong Granule water extract group was significantly lower than that in the Tougu Xiaotong Granule alcohol extract group (P < 0.05). Proliferation index and cyclin D1 mRNA and protein expression in the Tougu Xiaotong Granule water extract group were significantly higher than in the control group (P < 0.01), and these indices were significantly higher in the Tougu Xiaotong Granule water extract group than in the Tougu Xiaotong Granule alcohol extract group (P < 0.05). These findings suggest that Tougu Xiaotong Granule water/alcohol extract-containing serum promotes the proliferation of bone marrow stromal stem cells by upregulating Cyclin D1 expression and accelerating the progression of cell cycle; Tougu Xiaotong Granule water extract-containing serum exhibits better effects than Tougu Xiaotong Granule alcohol extract-containing serum. Related Articles | Metrics

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Magnesium affects the apoptosis and autophagy of CD4+ T cells in peripheral blood from healthy subjects Liang Rui-yun, Wu Wei, Huang Jin, Jiang Shan-ping 2012, 16 (41):  7723-7727.  doi: 10.3969/j.issn. 2095-4344.2012.41.025 Abstract ( 329 )   PDF (496KB) ( 488 )   Save BACKGROUND: Studies have demonstrated that magnesium (Mg) participates in the regulation of immune system including cell proliferation and apoptosis. Few studies have been reported regarding the effects of Mg concentration on the apoptosis and autography of CD4+ T lymphocytes in peripheral blood from healthy subjects. OBJECTIVE: To investigate the effects of Mg concentration on the apoptosis and autography of CD4+ T lymphocytes in peripheral blood from healthy subjects. METHODS: Peripheral blood CD4+T lymphocytes were isolated from 20 healthy subjects, and then cultured in low concentrations of Mg (0.2 and 0.4 mmol/L), physiological concentration of Mg (0.8 mmol/L) or high concentration of Mg (5 and 10 mmol/L). Apoptosis and autophagy levels were analyzed by flow cytometry after 1, 2, and 3 days. RESULTS AND CONCLUSION: The apoptosis and autophagy level of CD4+ T cells were significantly increased after cultured with low concentrations of extracellular Mg (0.2 and 0.4 mmol/L) (P < 0.05) and significantly decreased after cultured with high concentrations of extracellular Mg (5 and 10 mmol/L) (P < 0.05). Mg at a non-physiological concentration showed a concentration-dependent and time-dependent effect on apoptosis and autophagy of CD4+ T cells, but Mg at a physiological concentration (0.8 mmol/L) exhibited no effects on the apoptosis and autography of CD4+ T cells in the peripheral blood from healthy subjects (P > 0.05). Extracellular Mg deficiency exacerbated the ratio of apoptosis and autophgy in cultured peripheral CD4+ T cells from healthy subjects. Mg supplement reduced them and physiological concentration of Mg did not affect them. Related Articles | Metrics

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Construction and identification of recombinant adenovirus vectors encoding human transforming growth factor-beta 1 gene Zeng Zhao-xun, Yin Cheng-hui, Qiu Jun-qin, Chen Zong-xiong 2012, 16 (41):  7728-7732.  doi: 10.3969/j.issn.2095-4344.2012.41.026 Abstract ( 274 )   PDF (468KB) ( 475 )   Save BACKGROUND: Transforming growth factor-beta1 (TGF-β1) can promote the growth and proliferation of many cells and is one of main growth factors that regulate osteoblast differentiation of bone marrow mesenchymal stem cells. OBJECTIVE: To construct a recombinant adenoviral vector carrying the human TGF-β1 gene using the AdMax system which may be applicable in gene therapy of bone defect. METHODS: The hTGF-β1 gene sequence was amplified by PCR from cDNA template and then was inserted into shuttle plasmid pAV-MCMV-EGFP-3FLAG. The recombinant shuttle plasmid was constructed and transformed into E.coli DH5α.The recombinant pAV-MCMV-hTGF-β1 was obtained. RESULTS AND CONCLUSION: The recombinant pAV-MCMV-hTGF-β1 was packaged and amplified in 293 cells. Then the recombinant TGF-β1 adenovirus was constructed successfully. After measuring the titre of virus (1.25×1010 pfu/mL), the target gene was evaluated by PCR and Western Blot. These findings suggest that construction of hTGF-β1 adenoviral vector was successfully achieved using the AdMax system and may be available in the future study of TGF-β1 gene therapy. Related Articles | Metrics

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Expression of totipotency associated genes cPouV, cNanog and Sox2 in early chicken embryo Liu Ben-jie, Chen Zhi-sheng, Chen Sheng-feng, Ji Hui-qin, Wang Bing-yun, Deng Xian-bai 2012, 16 (41):  7733-7736.  doi: 10.3969/j.issn.2095-4344.2012.41.027 Abstract ( 261 )   PDF (390KB) ( 412 )   Save BACKGROUND: Studies have confirmed that totipotency associated genes cPouV, cNanog and Sox2 play an important role in regulating the pluripotency and self-renewal of chicken embryonic stem cells, but there lack studies on the expression and effect of the genes in early chicken embryo. OBJECTIVE: To detect the expression levels of cPouV, cNanog and Sox2 in early chicken embryo, and to find the effect of the genes in early chicken embryonic development. METHODS: 180 fresh fertilized eggs were divided into six groups, and took out after incubated for 0, 8, 16, 22, 32 and 42 hours. The embryo samples were obtained, and the real-time quantitative PCR was used to detect the expression levels of cPouV, cNanog and Sox2 with the internal control gene of β-actin. RESULTS AND CONCLUSION: The relative expressions of cNanog and cPouV have the same changes, the relative expressions of cNanog and cPouV were increased after incubated for 0, 8, 16 and 22 hours, and reached to peak after incubated for 22 hours and then decreased gradually. The relative expression of Sox2 showed a significant increasing trend. The totipotency associated genes of cPouV, cNanog and Sox2 have an important role in regulating the early embryonic development, but the effects are not exactly the same. Related Articles | Metrics

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Application prospect of bone marrow mesenchymal stem cells in the treatment of glioma Fan Cun-gang, Zhang Qing-jun 2012, 16 (41):  7737-7741.  doi: 10.3969/j.issn.2095-4344.2012.41.028 Abstract ( 261 )   PDF (607KB) ( 383 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) display extensive tropism for glioma and exert anti-tumor effect on glioma. OBJECTIVE: To investigate the therapeutic prospects of BMSCs in the treatment of glioma on the basis of reviewing related articles. METHODS: The first author retrieved CNKI database and PubMed database for concerning articles. The key words were “mesenchymal stem cells, bone marrow, glioma” in Chinese and English. A total of 51 articles were retrieved primarily by computer, and 29 articles were finally included for review according to inclusion criteria. RESULTS AND CONCLUSION: BMSCs have a strong migratory capacity towards glioma. In addition, BMSCs can inhibit the proliferation of glioma and induce the glioma cells differentiation. Moreover, the BMSCs carrying suicide genes, immune genes, tumor necrosis factor and oncolytic virus can exert stronger antitumor effect on glioma. BMSCs may serve as a promising vector for gene-targeted glioma therapy. Related Articles | Metrics

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Bone marrow mesenchymal stem cell transplantation combined with gene therapy for treatment of ischemic cerebrovascular disease Yan Shu-yang, Zhang Ji-zhi, Gao Bao-shan, Deng Dong-feng 2012, 16 (41):  7742-7746.  doi: 10.3969/j.issn.2095-4344.2012.41.029 Abstract ( 248 )   PDF (635KB) ( 400 )   Save BACKGROUND: Whether the transplanted bone marrow mesenchymal stem cells (BMSCs) can differentiate towards neural cells is regulated by cellular own genes and also depends on the influence of various signals in the external environment. OBJECTIVE: To review the biological characteristics of BMSCs, the theoretical evidence of transgenic therapy for treatment of ischemic cerebrovascular disease, and the research progress in animal experiments and clinical studies. METHODS: A computer-based online retrieval was performed by the first author in PubMed database to search the papers describing the biological characteristics of BMSCs, the theoretical evidence of transgenic therapy for treatment of ischemic cerebrovascular disease, and the research progress in animal experiments and clinical studies published in English or Chinese between January 2006 and December 2011, with the key words of bone marrow mesenchymal stem cells transplant, gene therapy, ischemic cerebrovascular disease (ICD). The repetitive studies and Meta analysis papers were excluded, and 25 papers were included in the final analysis. . RESULTS AND CONCLUSION: Cell transplantation and gene therapy for improving neurological function in treatment of ischemic cerebrovascular disease is an increasing research area. BMSCs are good vectors for cell gene engineering. After transgenic culture, BMSCs can be highly induced to differentiate into neural cells, which provide stem cells for treatment of ischemic cerebrovascular disease by cell transplantation. Great progress has been achieved in BMSCs transplantation combined with gene therapy for treatment of ischemic cerebrovascular disease. Nevertheless, its clinical application needs further investigation, in particular the problem of biological safety. Related Articles | Metrics

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Research advances in mechanisms by which bone marrow stem cells improve myocardial function Yu Gui-ping, Shen Zhen-ya 2012, 16 (41):  7747-7751.  doi: 10.3969/j.issn.2095-4344.2012.41.030 Abstract ( 211 )   PDF (268KB) ( 386 )   Save BACKGROUND: Bone marrow mesenchymal stem cells transplantation can significantly improve the cardiac structure reconfiguration and cardiac function through affecting the myocardial cell formation and myocardial angiogenesis. OBJECTIVE: To summarize the theoretical basis of bone marrow mesenchymal stem cells transplantation for the treatment of ischemic cardomyopathy through researching the effect of bone marrow mesenchymal stem cells transplantation on myocardial function. METHODS: The Chinese Biomedical Literature database and Medline database from 1994 to 2011 were used to search the reviews and reports that relate to the application of bone marrow mesenchymal stem cells transplantation for the treatment of ischemic cardomyopathy, and the research progress was analyzed. RESULTS AND CONCLUSION: A total of 37 articles were included for the review. Autologous application of bone marrow mesenchmal stem cells transplantation can be amplified in vitro without immune rejection and can avoid the ethical controversy. Research of bone marrow stem cells for the treatment of acute myocardial infarction is still in the early stage. With the development of the research, bone marrow mesenchymal stem cells become a new method for the treatment of ischemic cardiomyopathy if we can find an effective and safe dosage. Related Articles | Metrics

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Application of endothelial progenitor cells in the treatment of ischemic cerebrovascular disease Zhao Rui, Huang Qing-hai, Zhao Wen-yuan, Hong Bo, Xu Yi, Liu Jian-min 2012, 16 (41):  7752-7761.  doi: 10.3969/j.issn.2095-4344.2012.41.031 Abstract ( 314 )   PDF (737KB) ( 438 )   Save BACKGROUND: Previous studies have found that bone marrow and peripheral blood endothelial progenitor cells play an important role in angiogenesis in the adult under pathological conditions. There was the formation of new blood vessels in the ischemic area after cerebral infarction, so endothelial progenitor cells were also involved in this process. The increase of endothelial progenitor cells numbers, as well as the maximum degree of participation in the organization of new blood vessels, provides an important way for the treatment of cerebrovascular disease. OBJECTIVE: To multivariately analyze the literature on the application of endothelial progenitor cells in the treatment of cerebrovascular disease through SCI database and CNKI database in order to understand the trend of endothelial progenitor cells in the treatment of cerebrovascular disease, the characteristics of endothelial progenitor cells, and the role of data analysis and its application in patients with cerebrovascular disease progress. METHODS: A retrieval was performed for the literature of the application of endothelial progenitor cells in the treatment of cerebrovascular disease, using key words “endothelial progenitor cells, precursor cell, stroke, brain ischemia or cerebral ischemia, cerebral infarction, ischemic cerebral vascuIar disease” during 2002-01 and 2011-12 in Scopus database and CNKI database. The retrieval results were analyzed, and the trends were described in words and graphics. RESULTS AND CONCLUSION: Endothelial progenitor cells are bone marrow precursor cells in adults, which can migrate to the peripheral blood and differentiate into mature endothelial cells. They have proliferative potential and can participate in the vascular endothelial repair and new blood vessel formation in ischemic organs. According to the biological properties and role of endothelial progenitor cells, the present study found that endothelial progenitor cells can prevent cerebral artery atherosclerosis, and angiogenesis after focal cerebral ischemia, the forecast evaluation of cerebral ischemic disease, and cerebral hemorrhage after treatment. Thus, endothelial progenitor cells in the treatment of cerebrovascular disease show a wide range of applications. Related Articles | Metrics

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Research and application of seeded cells in reconstruction of jawbone defects Zou Wei, Liu Hui-tong, Li Teng, Wang Min 2012, 16 (41):  7762-7767.  doi: 10.3969/j.issn.2095-4344.2012.41.032 Abstract ( 299 )   PDF (624KB) ( 426 )   Save BACKGROUND: The morphology as well as the establishment of occlusal relationship should be considered during the construction of jawbone defects. OBJECTIVE: To summarize the research status of seeded cells of bone tissue engineering applied to the treatment of jawbone defects in recent years. METHODS: A computer-based online search was conducted in PubMed database and CNKI database for the relate articles published from June 1994 to April 2011 with the key words “bone tissue engineering, seeded cells, osteogenic differentiation” in Chinese and English. The irrelevant or repetitive articles were eliminated by reading the titles and abstracts. A total of 58 articles were selected to review. RESULTS AND CONCLUSION: This paper has comprehensively analyzed the current research of various seeded cells used in bone tissue engineering, bone marrow stromal stem cells are most studied, and the research on umbilical cord blood mesenchymal stem cells and umbilical cord mescenchymal stem cells is also popular as scholars can overcome the application disadvantages of mesenchymal stem cells. Embryonic stem cells are expected to become the important seeded cells in bone tissue engineering for repair of jawbone defects if there are no ethical problems. Genetically modified seed cells can enhance the proliferation and directed osteogenic differentiation capacity of seeded cells in bone tissue engineering. But genetic engineering technique is still needed to be improved, and its safety and effectiveness need further discussion. Related Articles | Metrics

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Role of periodontal ligament stem cells during regeneration of periodontal tissue Yu Hai-tao, Maimaitili·Gulibaha 2012, 16 (41):  7768-7771.  doi: 10.3969/j.issn.2095-4344.2012.41.033 Abstract ( 953 )   PDF (469KB) ( 688 )   Save BACKGROUND: It is difficult to regenerate the periodontal tissue because the number of stem cells is not enough and their biological function is insufficient. OBJECTIVE: To summarize the progress and perspective of periodontal ligament stem cells for the periodontal tissue regeneration in dentistry. METHODS: The first author searched PubMed database and CNKI database (2000-01/2012-07) for articles regarding isolation, identification, and relevant factors of periodontal ligament stem cells published between January 2000 and July 2012 using the key words of “periodontal ligament stem cells” in English and “periodontal ligament, stem cells” in Chinese. Repetitive articles were excluded, and 26 articles were included in the final analysis. RESULTS AND CONCLUSION: Using tissue engineering technology, periodontal ligament stem cells were used as the seed cells of periodontal tissue regeneration and transplanted into the defected area after in vitro culture and amplification. Results showed that periodontal ligament stem cells can promote the regeneration of periodontal attachment tissue and shorten the healing period. The study of periodontal ligament stem cells has become a key point in the field of tooth tissue engineering. Related Articles | Metrics

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Neural differentiation capacity and potential clinical application of adipose-derived mesenchymal stem cells Han Chao, Song Lin, Zou Wei, Liu Jing 2012, 16 (41):  7772-7776.  doi: 10.3969/j.issn.2095-4344.2012.41.034 Abstract ( 371 )   PDF (537KB) ( 374 )   Save BACKGROUND: It has been recently confirmed that adipose-derived stem cells have the characteristics of the mesenchymal stem cells (MSC) residing in bone marrow. They have the same cell surface markers and are capable of differentiating into the same cell types, including osteoblasts, chondrocytes, myoblasts, adipocytes, and neuron-like cells. OBJECTIVE: To review the biological characteristics of adipose-derived mesenchymal stem cells and their clinical application for treatment of somatic nervous system diseases. METHODS: A computer-based online retrieval of PubMed database was performed to search the papers describing neural differentiation capacity of adipose-derived mesenchymal stem cells and their clinical application for treatment of nervous system diseases published during January 1989 to January 2012 using the key words mesenchymal stem cell, nervous system disease, stem cell therapy. A total of 733 papers were retrieved, and 26 were included for final analysis. RESULTS AND CONCLUSION: Adipose tissue is ubiquitous in human body, can be easily harvested and is of high purity, with high activity and strong amplification, which make adipose-derived mesenchymal stem cells to become the ideal seed cells for clinical application in regeneration medicine. Recent studies have pointed out that adipose-derived mesenchymal stem cells greatly promote the functional recovery of injured nerve. Nevertheless, the mechanism underlying curative effects of adipose-derived mesenchymal stem cells are still in the exploratory stage, and the efficacy and safety of adipose-derived mesenchymal stem cells in treatment of nervous system disease need to be investigated by a large number of clinical case data. All these restrict the clinical application of adipose-derived mesenchymal stem cells. Related Articles | Metrics

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Umbilical cord blood stem cell transplantation combined with mouse neural growth factor application and physical rehabilitation therapy for infantile cerebral palsy Xing Li-he, Zhang Li-xin, Zhang Li-li, Sun Li-fang, Dong Yan-hui, Liu Ying, Guo Li-jie 2012, 16 (41):  7777-7781.  doi: 10.3969/j.issn.2095-4344.2012.41.035 Abstract ( 262 )   PDF (452KB) ( 580 )   Save BACKGROUND: Umbilical cord blood stem cell transplantation is an optimal therapeutic method of nervous system diseases. OBJECTIVE: To investigate the clinical efficacy of umbilical cord blood stem cell transplantation combined with mouse neural stem cell application and physical rehabilitation therapy for infantile cerebral palsy. METHODS: 90 patients with cerebral palsy were randomly and evenly divided into three groups: umbilical cord blood stem cells group (umbilical cord blood mesenchymal stem cells + mouse neural growth factor + physical rehabilitation therapy), routine treatment group (mouse neural growth factor + physical rehabilitation therapy) and control group (no effective treatment). RESULTS AND CONCLUSION: Prior to treatment, there was no significant difference in gross motor function measure (GMFM) scores of each functional item among three groups (P > 0.05). Compared with prior to treatment, at 3 months after treatment, the GMFM scores of A, B, D items were significantly increased in the umbilical cord blood stem cells group and routine treatment group (P < 0.05); GMFM scores of A, B functional items were significantly higher in the umbilical cord blood stem cells group than in the routine treatment group (P < 0.05); there was no significant difference in GMFM scores of each functional item in the control group (P > 0.05). These results showed that umbilical cord blood mesenchymal stem cell transplantation combined with mouse neural stem cell application and physical rehabilitation therapy exhibits better curative effects in treatment of infantile cerebral palsy than mouse neural growth factor application combined with physical rehabilitation therapy. Related Articles | Metrics

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Autologous peripheral blood stem cell in combination with bone marrow-derived mesenchymal stem cell transplantation for two patients with non-Hodgkin lymphoma Luo Zi-mian, Zhou Xin-fu, Kuang Wen-yong, Lei Xiao-yu, Tan Zhen-qing, Yin Ya-fei, Tang Tie-gang, Liu Li-hua, Liu Kang, Li Jia-yuan 2012, 16 (41):  7782-7786.  doi: 10.3969/j.issn.2095-4344.2012.41.036 Abstract ( 353 )   PDF (380KB) ( 396 )   Save BACKGROUND: Peripheral blood stem cell (PBSC) transplantation in combination with bone marrow-derived mesenchymal stem cell (BMSC) transplantation may minimize hematopoietic reconstruction delay or failure caused by hemopoietic microenvironment damage. OBJECTIVE: To investigate the safety and effect of BMSC+ PBSC transplantation in the treatment of non-Hodgkin lymphoma. METHODS: Two patients diagnosed as non-Hodgkin lymphoma were treated with chemotherapy for 5 or 6 cycles using R-CHOP protocols (rituximab, cyclophosphamide, Oncovin, and prednisone). BMSCs from autologous bone marrow were cultured prior to autologous PBSC mobilization using cyclophosphamide, granulocyte colony-stimulating factor or rituximab, cyclophosphamide, and granulocyte colony-stimulating factor. Preconditioning protocols used rituximab, cyclophosphamide, etoposide or rituximab, cyclophosphamide, liposome adriamycin, and dexamethasone. Mononuclear cells of 2.98×106/kg, 3.84×108/kg and BMSCs of 3.8×106/kg, 3.96×106/kg were infused. RESULTS AND CONCLUSION: In case 1, the white blood cells decreased to the lowest level, 0.1×109/L, neutrophil, 0×109/L, 10 days following transplantation, and the platelet reduced to the minimal level of 45×109/L up to 12 days. Peripheral blood restored to normal at 15 days after transplantation. The white blood cells and platelet reduced to the lowest levels up to 5 days following transplantation in case 2 and the peripheral blood restored to normal at 9 days. Transplantation-related complications included acute upper respiratory tract infection and external hemorrhoid infection, which were controlled by treatment. Results indicated that in the treatment of malignant lymphoma, autologous BMSC+ PBSC transplantation rapidly reconstructed hematopoiesis, and the tumor or swollen lymph node disappeared. The short-term effects were evident, but the long-term effect requires further investigation. Related Articles | Metrics

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Transplantation of umbilical cord mononuclear cells for treatment of infantile spastic cerebral palsy Xu Rong, Liu Bo, Duan Da, Fan Meng-yao, Liu Bin, Teng Xiao-hua, Zhao Zhen-yu, Lu Ming 2012, 16 (41):  7787-7790.  doi: 10.3969/j.issn.2095-4344.2012.41.037 Abstract ( 230 )   PDF (437KB) ( 379 )   Save BACKGROUND: A large number of studies have demonstrated that umbilical cord mesenchymal stem cells cultured in vitro can express the surface markers of neurons and glial cells under induction and exhibit replacing and repairing effects for nervous system disease. OBJECTIVE: To study the therapeutic effect and safety of transplantation of human umbilical cord blood mononuclear cells in treatment of infantile spastic cerebral palsy. METHODS: Umbilical cord blood was harvested from full term deliveries scheduled for cesarean section and anti-coagulated with heparin. Umbilical cord blood mononuclear cells were isolated by lymphocyte separation medium, purified by Nash differential adhesion, and then injected into subarachnoid cavity of 30 infantile patients with spastic cerebral palsy. RESULTS AND CONCLUSION: Compared with prior to treatment, the muscle spastic and joint range of motion significantly improved after treatment (P < 0.05). There were no adverse events and complications after transplantation and during follow-up period. Related Articles | Metrics