Table of Content

02 April 2010, Volume 14 Issue 14 Previous Issue    Next Issue

For Selected:

Download Citations EndNote Reference Manager ProCite BibTeX RefWorks

Toggle Thumbnails

Select

Differentiation of rhesus monkey mesenchymal stem cells into neuron-like cells by sonic hedgehog factor Song Ge, Zhang Yang, Liu Bing-qian, Zheng Wei-wei, Sun Xue-rong 2010, 14 (14):  2471-2475.  doi: 10.3969/j.issn.1673-8225.2010.14.001 Abstract ( 120 )   PDF (1504KB) ( 410 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) differentiating into neural cells is an effective way of cell therapy of nervous system disease. However, the methods used nowadays still need to be improved. OBJECTIVE: To induce the differentiation of rhesus monkey BMSCs into neuron-like cells by using sonic hedgehog factor. METHODS: Rhesus monkey BMSCs differentiating into neuron-like cells was induced by typical retinoic acid and sonic hedgehog factor. Rhesus monkey BMSCs were isolated and cultured by density gradient centrifugation method. Cell growth was observed under an inverted phase contrast microscope and cell growth curve was determined using MTT assay. Flow cytometry was performed to characterize the phenotype of BMSCs, and immunohistochemistry was utilized to assess differentiated cells. Ultra-structure of the differentiated cells was observed by transmission and scanning electron microscopes. RESULTS AND CONCLUSION: Rhesus monkey BMSCs cultured in vitro were identified by flow cytometry, with high homogenicity. Following sonic hedgehog factor disposal for 7 days, differentiated cells were mainly positive for neurone specific enolase, neurofilament protein, Tau and glial fibrillary acidic protein (GFAP). Image statistical analysis found that in sonic hedgehog factor scheme, neural stem cells marker Nestin positive rate was significantly higher compared with the retinoic acid scheme (P < 0.01). GFAP-positive rate was greater in the retinoic acid scheme than in the sonic hedgehog factor scheme (P < 0.05). Results indicated that sonic hedgehog factor scheme is an effective pathway of rhesus monkey BMSC differentiation into neuron-like cells. Related Articles | Metrics

Select

Differentiation of rat bone marrow mesenchymal stem cells into neuron-like cells in vitro: Induction of Wnt3a signaling molecules Yan Wen-zhu, Qin Shu-jian, Liu Xue-zheng, Li De-hua 2010, 14 (14):  2476-2480.  doi: 10.3969/j.issn.1673-8225.2010.14.002 　 Abstract ( 84 )   PDF (1279KB) ( 509 )   Save BACKGROUND: Wnt signaling pathway is the key to regulation of cell proliferation and differentiation. The evidence suggests that this pathway participates in the neural precursor cell proliferation, differentiation and determination of the regulation of cell fate. At present, the Wnt signaling pathway effects on the mesenchymal stem cells (MSCs) differentiated into neuron-like cells have not been reported. OBJECTIVE: To seek Wnt signal molecule that promotes the differentiation of MSCs into neuron-like cells. METHODS: MSCs were isolated from the femur of Sprague Dawley rats in vitro using the density gradient centrifugation, and then cultured. Following passage, cell surface markers CD29, CD44, CD34 and CD45 were measured using morphology and flow cytometry. These cells were selected and evaluated. Using neurotrophic factor and basic fibroblast growth factor combined with Wnt3a and Wnt5a induction scheme, effects of Wnt3a and Wnt5a during the differentiation of MSCs into neuron-like cells were compared utilizing immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR). Simple culture of basic fibroblast growth factor served as controls. RESULTS AND CONCLUSION: Following culture and passage of MSCs, cells adhered to the wall, showing even spindle shape. Flow cytometry showed great expression of CD29 and CD44 and low expression of CD34 and CD45. Following Wnt3a induction, cells were positive for nestin, neuron specific enolase, but no significantly expressed glial fibrillary acidic protein. Following induction, cell viability was good. In the Wnt5a induction and control groups, weakly positive expression of nestin was found, but neuron specific enolase and glial fibrillary acidic protein were negative. RT-PCR results demonstrated that nestin expression was detected in the Wnt3a induction group before and after induction. Significant amplified bands for neuron specific enolase were detected at day 5 following induction, and became more significant at day 10. Weak bands for glial fibrillary acidic protein were visible at day 10 following induction. These indicated that Wnt3a can promote the differentiation of MSCs into neuron-like cells in vitro. Related Articles | Metrics

Select

Induction and differentiation of rat bone marrow mesenchymal stem cells into neuron-like cells in vitro Gu Yan-jiao, Li Bo, Gao Zhi-an 2010, 14 (14):  2481-2484.  doi: 10.3969/j.issn.1673-8225.2010.14.003　 Abstract ( 116 )   PDF (1147KB) ( 359 )   Save BACKGROUND: It is a key to choose an appropriate method to trans-differentiated bone marrow mesenchymal stem cells (BMSCs) into neuron-like cells for clinical therapy of neural system injury. OBJECTIVE: To observe the feasibility of the differentiation of rat BMSCs supplemented with rat dentate gyrus extract (DGE) into neuron-like cells. METHODS: DGE was applied to induce rat BMSC trans-differentiation, using 20, 40, 60, 80 mg/L protein concentration. Neuron differentiation was measured using Western blot to screen an optimal dosage. After trans-differentiation, different concentration treated cells were collected. Morphology change was observed following differentiation under an inverted microscope. Neuron specific enolase (NSE) and NeuN expression was determined using Western blot. NeuN expression in cells was detected using immunofluorescence technique. RESULTS AND CONCLUSION: After 7 days, 60 mg/L DGE was the optimal induction dose detected by Western blot. With increased concentration, NSE and NeuN expression was increased. At 80 mg/L mass concentration, NSE and NeuN expression was reduced. At 60 mg/L DGE, BMSCs following induction became long, with synapses. Immunofluorescence NeuN staining found that neuron-like cells were positive for NeuN following induction. Results indicated that DGE is an effective biological inductor that can induce BMSC differentiation. Related Articles | Metrics

Select

Immunomodulatory effect of Wharton’s jelly-derived mesenchymal stem cells from human umbilical cord on human peripheral blood T lymphocytes   Zhou Chang-hui, Tian Yi, Yang Bo, Hu Xiang, Jiao Hong-liang, Zhou Yun-fan, Wang Cheng-chun, Gu Chen-xi,Lei Ning-jing, Guan Fang-xia 2010, 14 (14):  2485-2491.  doi: 10.3969/j.issn.1673-8225.2010.14.004 Abstract ( 124 )   PDF (2168KB) ( 384 )   Save BACKGROUND: Bone marrow mesenchymal stem cells have low immunogenicity and immunomodulatory effect, but there are seldom reports concerning the immunomodulatory effect of Wharton’s jelly-derived mesenchymal stem cells of human umbilical cord and its mechanims. OBJECTIVE: To investigate the immunomodulatory effects and mechanisms of Wharton’s jelly-derived mesenchymal stem cells of human umbilical cord on varient peripheral blood T lymphocytes. METHODS: Mesenchymal stem cells were isolateded from Wharton’s jelly of human umbilical cord by tissue culture. T lymphocytes from human peripheral blood were stimulated by phytohemagglutinin and co-cultured with umbilical cord Wharton’s jelly-derived mesenchymal stem cells and umbilical cord Wharton’s jelly-derived mesenchymal stem cells supernatant respectively to measure A value following 72 hours of coculture using multifunctional microplate reader. Expression of cytokines including transforming growth factor-beta 1 (TGF-β1) and interferon-γ (IFN-γ) was evaluated by enzyme-labeled immunosorbent assay. RESULTS AND CONCLUSION: Wharton’s jelly-derived mesenchymal stem cells could inhibite the proliferation of T lymphocytes induced by phytohemagglutinin. The proliferation inhibition rate was 56% (P < 0.01). Wharton’s jelly-derived mesenchymal stem cells supernatant also had inhibitory effects on proliferation of T lymphocytes induced by phytohemagglutinin, in a dose-dependent fashion. The proliferation inhibition rates were 8.3% and 27% respectively in the 50% Wharton’s jelly-derived mesenchymal stem cells supernatant and 100% Wharton’s jelly-derived mesenchymal stem cells supernatant groups (P < 0.05). Wharton’s jelly-derived mesenchymal stem cells significantly decreased γ-interferon secrted from T-lymphocytes (P < 0.05). The secretion of TGF-β1 was lower in the coculture of Wharton’s jelly-derived mesenchymal stem cells and T lymphocytes group than Wharton’s jelly-derived mesenchymal stem cells alone group (P < 0.05). These indicated that Wharton’s jelly-derived mesenchymal stem cells and Wharton’s jelly-derived mesenchymal stem cells supernatant have inhibitory effects on proliferation of T lymphocytes induced by phytohemagglutinin. The mechanims may be associated with cell contant and inhibition of γ-interferon secrted from T-lymphocytes. Related Articles | Metrics

Select

Isolation of human umbilical cord mesenchymal stem cells and differentiation into adipocytes and osteblasts   He Shao-qing, Luo Zhen-yu, Liu Qiu-ying, Zhou Xiang-rong, Deng Ming-quan, Luo Xin, Yao Run-si, Gao Zhi, Wang Yi-fei 2010, 14 (14):  2492-2496.  doi: 10.3969/j.issn.1673-8225.2010.14.005 Abstract ( 150 )   PDF (1508KB) ( 477 )   Save BACKGROUND: Culture condition, isolation method and efficiency are different in reported human umbilical cord-derived mesenchymal stem cells, which lack of unified identification standards. Therefore, it is necessary to establish a high-efficiency and economical culture system for human umbilical cord-derived mesenchymal stem cells (hUCMSCs). OBJECTIVE: To isolate hUCMSCs and induced differentiate into adipocytes and osteblasts. METHODS: The hUCMSCs were isolated form human umbilical cord by tissue adherence and digested with collagenase. The morphology, proliferation and immunophenotype of the 3rd passage cells were analyzed, and then cells were induced to osteogenic and adipogenic differentiation in vitro. RESULTS AND CONCLUSION: The hUCMSCs isolated from human umbilical cord by tissue adherence and digested with collagenase could be cultured and proliferated in vitro. Flow cytometry analysis revealed that the hUCMSCs were positive for CD29, CD44, CD59, CD105, but were negative for CD40, CD86 and HLA-DR. These cells could be induced to differentiate into adipocytes and osteblasts under proper inducing conditions. The hUCMSCs retained the appearance and phenotype even after being expanded more than 40 passages in vitro. This confirmed that the existence of MSCs in human umbilical cord and they had the capacity of differentiating into adipocytes and osteblasts. Related Articles | Metrics

Select

Effects of different serum microenvironments on culture of rat bone marrow mesenchymal stem cells in vitro   Zheng Jing-hui, Li Yong-hua, Wang Li-ping, Jian Wei-xiong, Huang Xian-ping, Yuan Zhao-kai 2010, 14 (14):  2497-2502.  doi: 10.3969/j.issn.1673-8225.2010.14.006 Abstract ( 212 )   PDF (657KB) ( 484 )   Save BACKGROUND: During culture of bone marrow mesenchymal stem cells (BMSCs), a certain serum is commonly added in the basic medium, such as calf serum and fetal bovine serum, but there are potential biological safety risks. OBJECTIVE: To study the effects of different serum microenvironments on in vitro culture of rat BMSCs. METHODS: BMSCs were harvested from adult rat bone marrow, and cultured in vitro by whole bone marrow adherence method. The cells were cultured under the following serum microenvironment. The primary cells of autoserum group were cultured with autoserum, changing the medium with fetal bovine serum after passage. The primary cells of homogeneity foreign serum group were cultured with homogeneity foreign serum, changing the medium with fetal bovine serum after passage. The primary cells of fetal bovine serum group were cultured with fetal bovine serum, and cultured with fetal bovine serum after passage. The primary cells of Dulbecco's modified Eagle's medium (DMEM) group were cultured with serum-free DMEM, changing the medium with fetal bovine serum after passage. The morphologic changes in BMSCs were detected under an inverted phase contrast microscope. Attachment rate and growth curve were measured. Surface marker CD11b, CD45 and CD90 expression was analyzed by flow cytometry. RESULTS AND CONCLUSION: In autoserum and homogeneity foreign serum groups, the homogenicity and degrees of fusion of cell morphology were improved in comparison with other two groups and the day of first passage was less than other groups. The attachment rate was greater in the autoserum, homogeneity foreign serum and fetal bovine serum groups than the DMEM group at 24, 48, 72 hours (P < 0.01). Doubling rate was fastest in the growth curve of autoserum group, followed by homogeneity foreign serum group and fetal bovine serum group. However, no doubling phenomenon was found in the DMEM group. Flow cytometry results demonstrated that the rates of CD11b-positive and CD45-positive cells at passage 3 were above 98% under medium containing serum, and CD90-positive rate was less than 2%. We could obtain BMSCs of higher purity. However, CD11b-positive rate was 95.83%, CD90-positive rate was 2.07%, but CD45 positive rate was only 64.79% under serum-free microenvironment. BMSC purity was significantly lower under serum-free microenvironment than under serum microenvironment. Results indicated that the microenvironment of rat autoserum can improve the attachment rate, growth and purification of BMSCs. Related Articles | Metrics

Select

In vitro differentiation of human bone marrow mesenchymal stem cells into hepatocyte-like cells: Effect of hepatocyte growth factor and epidermal growth factor Xiong Jian-yong, Chen Bin, Ni Yong 2010, 14 (14):  2503-2507.  doi: 10.3969/j.issn.1673-8225.2010.14.007 Abstract ( 128 )   PDF (643KB) ( 365 )   Save BACKGROUND: Previous research has demonstrated human bone marrow mesenchymal stem cells (HMSCs) differentiate into hepatocyte-like cells; however, biological characteristics and differentiation mechanism remain unclear, and differentiation system remains immature. OBJECTIVE: To investigate the feasibility of hepatocyte growth factor (HGF) and epidermal growth factor (EGF) to induce the differentiation of HMSCs into hepatocyte-like cells. METHODS: HMSCs were obtained from patients with esophageal cancer and were separated by density gradient centrifugation combined with attachment method. The phenotypes of MSCs were identified by flow cytometry. The third-passage HMSCs were divided into four groups: HGF (adding 20 µg/L HGF), EGF (adding 20 µg/L EGF), HGF + EGF, and blank control groups. Morphology was observed using inverted microscope. At days 7 and 14 after induction, α-fetoprotein and albumin mRNA expressions were detected using RT-PCR assay. RESULTS AND CONCLUSION: The HMSCs did not express hematopoietic cell CD34 and CD35, but strongly expressed β1-integrin CD29 and matrix receptor CD44. HMSCs changed from long fusiform shape to polygon or similar round shape in the HGF, EGF, and HGF + EGF groups. At days 7 and 14 after induction, α-fetoprotein and albumin mRNA expressions were positive. However, polygon cells were not observed in the blank control group, and α-fetoprotein and albumin mRNA expressions were negative. This suggested that HGF, EGF, and HGF + EGF could induce the differentiation of HMSCs into hepatocyte-like cells; however, their differentiation ability still needs to be further semi-quantitatively analyzed using immunohistochemical staining. Related Articles | Metrics

Select

Rat bone marrow mesenchymal stem cells isolated and purified by whole bone marrow adherent method and differential passage: A comparison with density gradient centrifugation Lin Chu-wei, Zhou Sheng-hua, Du You-you 2010, 14 (14):  2508-2512.  doi: 10.3969/j.issn.1673-8225.2010.14.008 Abstract ( 148 )   PDF (1308KB) ( 612 )   Save BACKGROUND: At present, there are two main methods of isolating and purifying bone marrow mesenchymal stem cells (BMSCs): density gradient centrifugation and the whole bone marrow adherent method. The former has complicated procedures, and the latter has simple operation, but the purified outcomes are not ideal. OBJECTIVE: To establish a rat BMSCs isolated and cultured in vitro purification methods on the basis of the whole bone marrow adherence and isolation of BMSCs in combination of differential passage digestion method. METHODS: By whole bone marrow adherent culturing to isolate and differential digestion and passage of rat BMSCs, the speed of MSCs in the process of digestion and passage was quicker than other bone marrow cells, as well as the characteristics of adherent speed, instead of density gradient centrifugation to separate and purify MSCs, and their morphological characteristics were observed. Cell growth and proliferation of two kinds of culture method were compared with the density gradient centrifugation separation. Alkaline phosphatase and oil red staining results were observed to verify BMSC differentiation capacity, to detect the cell surface markers, to validate immune properties and to test its purity. RESULTS AND CONCLUSION: The whole bone marrow adherent culture method isolated and differentially subcultured rat BMSCs. Flow cytometry and osteogenic and adipogenic culture results displayed cytoimmunity property, purity and differential capacity, which have no significant difference compared with density gradient centrifugation. However, cell viability and reproductive activity are obviously elevated. Related Articles | Metrics

Select

Superparamagnetic iron oxide and chlormethylbenzamido-1, 1'-dioctadecyl-3, 3, 3', 3'-tetramethylindocarbocyanine labeling and tracing bone marrow mesenchymal stem cells  Dou Xing-kui, Guo Tao, Si Yong-yu, Wan Lin-jun, Huang Qing-qing, Kang Bo 2010, 14 (14):  2513-2517.  doi: 10.3969/j.issn.1673-8225.2010.14.009 Abstract ( 76 )   PDF (1379KB) ( 434 )   Save BACKGROUND: The success of cell therapy will depend on the ability to monitor the fate of transplanted cells in vivo. Superparamagnetic iron oxide (SPIO) labeling is an ideal magnetic resonance contrast medium, and it offers the potential for non-invasive tracking of implanted cells. The chlormethylbenzamido-1,1' - dioctadecyl- 3, 3, 3',3' - tetramethylindocarbocyanine (CM-DiI) labeling does not have cytotoxicity, so it cannot influence cell growth. OBJECTIVE: To investigate the effects of SPIO and CM-DiI labeling and tracking on bone marrow mesenchymal stem cells (BMSCs). METHODS: Porcine BMSCs were isolated and cultured by the whole bone marrow method. BMSCs were labeled with SPIO containing 50 mg/L Ferrum and CM-DiI. The labeled BMSCs were transplanted into porcine myocardial infarction model via intracoronary infusion. The frozen sections of the cardiac tissues were obtained after 4 weeks. RESULTS AND CONCLUSION: Efficiency of SPIO and CM-DiI labeling BMSCs was nearly 100%. The SPIO and CM-DiI labeled BMSCs could be found in the cardiac muscle tissues at 4 weeks after transplantation. SPIO and CM-DiI labeling BMSCs were efficiently tracked in vivo. Related Articles | Metrics

Select

Expression of pDsRed-C1-CDNF eukaryotic expression vector in rat bone marrow mesenchymal stem cells Zhang Jun, Niu Chao-shi, Gao Ge, Tang Shen-feng, Li Jing 2010, 14 (14):  2518-2522.  doi: 10.3969/j.issn.1673-8225.2010.14.010　 Abstract ( 136 )   PDF (1388KB) ( 361 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (MSCs) are a kind of adult stem cells with multi-potential differentiation property. At present, it has served as cell carrier for the treatment of Parkinson's disease. OBJECTIVE: To construct pDsRed-C1-CDNF eukaryotic expression vector and induce its expression in rat MSCs. METHODS: CDNF gene was amplified from mouse tissues using RT-PCR, and sequence with Xho I, BamHI restriction enzyme cutting site. The CDNF gene was inserted into the eukaryotic expression vector pDsRed-C1 encoding red fluorescent protein gene. The plasmid pDsRed-C1-CDNF was constructed and transfected into rat bone marrow MSCs. RESULES AND CONCLUSION: The pDsRed -C1-CDNF recombinant plasmid was confirmed by double digestion of Xho I and BamHI restriction enzyme or single digestion of BamHI, and PCR sequence. Results show that the recombinant pDsRed-C1-CDNF eukaryotic expression vector was successfully constructed. Related Articles | Metrics

Select

Effect of hyperbaric oxygen on differentiation and Wnt3 expression of bone marrow mesenchymal stem cells Chen Chong-feng, Yang Yu-jia, Yao Yue, Wang Qing-hong, Li Meng 2010, 14 (14):  2523-2527.  doi: 10.3969/j.issn.1673-8225.2010.14.011 Abstract ( 94 )   PDF (1474KB) ( 358 )   Save BACKGROUND: Hyperbaric oxygen (HBO) treatment promotes the proliferation and differentiation of endogenous neural stem cells in neonatal rats following hypoxic/ischemic brain damage (HIBD). The Wnt signaling pathway is associated with neurogenesis. However, there are few data recording the role of HBO in the differentiation of neural stem cells in vitro. OBJECTIVE: To observe the effect of HBO on differentiation and Wnt3 expression of bone marrow mesenchymal stem cells (BMSCs). METHODS: BMSCs were isolated and cultured. The rat BMSCs of passages 3-5 were cultured in DMEM/F12 (1: 1) medium with basic fibroblast growth factor, epidermal growth factor and B27 for 24 hours. The induced BMSCs were randomly divided into two groups: control group (no treatment) and HBO group (HBO, 0.10 MPa, 60 minutes stabilizing pressure with at least 90% oxygen). The neuron specific encloase (NSE), glial fibrillary acidic protein (GFAP) and O4 marked oligodendrocyte immunocytochemistry were detected by immunofluorescent staining, and Wnt3 protein expression was detected by Western-blot.    RESULTS AND CONCLUSION: BMSCs cultured in classic medium of neural stem cells could significantly induce the expression of nestin. The expression of NSE and O4 of HBO group was greater than control group (P < 0.01), but GFAP expression displayed no significant difference between the groups (P > 0.05). Western blot showed HBO could enhance the Wnt3 expression (P < 0.05). Results show that HBO can induce BMSCs to differentiate into neural cells and oligodendrocyte, which is correlated with the activation of the Wnt3 protein. Related Articles | Metrics

Select

Oriented migration of intravenously administrated mesenchymal stem cells transfected with adenovirus vector mediated green fluorescence protein in the lung tissues of pulmonary emphysema rats   Sun Yan-wei, Li Bao-ping, Wang Xuan, Zhang Lei, Lu Peng-yan, Zhao Xiao-yan, Guo Zi-kuan 2010, 14 (14):  2528-2532.  doi: 10.3969/j.issn.1673-8225.2014.14.012 Abstract ( 130 )   PDF (1411KB) ( 478 )   Save BACKGROUND: Recently, application of stem cells and growth factor to promoting lung regeneration in repair of emphysema lesion has been a hot focus in study. Thus, it is worth to pay attention on whether stem cells carrying relevant foreign growth factor gene can repair emphysema lesion. OBJECTIVE: To evaluate the efficiency of adenovirus vector mediated green fluorescence protein (Ad-GFP) transfecting bone marrow mesenchymal stem cells (BMSCs) and its effect on the cell proliferation, to explore oriented migration of intravenously administrated BMSCs transfected with Ad-GFP in the lung tissues of pulmonary emphysema rats. METHODS: MSCs were separated and purified from the bone marrow of rats by density gradient centrifugation and by adherence. At different multiplicity of infection (MOI), transfection efficiency was observed by laser confocal microscopy. At 48 hours of transfection, MTT method was used to evaluate the proliferation of MSCs. A total of 16 Wistar rats were randomly divided into emphysema model group and control group (n = 8). Model rats were established by exposure to cigarette smoke. MSCs, transfected with Ad-GFP, were grafted into the body of rats via tail vein. Lungs derived at 24 hours after implantation, and frozen sections were made. Migration and survival of MSCs in the lung tissues were observed by fluorescence microscopy. RESULTS AND CONCLUSION: MSCs from Wistar rats were successfully cultured, grew well and infected by Ad-GFP. The highest transfection effincincy (88.42 %) could be achieved at MOI of 200. Green fluorescent protein labeling had little effect on proliferation of MSCs by different MOI (P > 0. 05). At 24 hours posttransplantation, the green fluorescence-positive tissue was found in the lung tissues of emphysema model group and control group. Compared with control group, the expression of GFP in lung tissues was higher in emphysema model group (P < 0. 05). These suggested that introduction of target gene cannot affect proliferation and homing property of BMSCs. Related Articles | Metrics

Select

Autologous activated Schwann cells transplantation via subarachnoid space for rat spinal cord injury  Wang Chun-yuan, Feng Shi-qing, Liu Yang, Han Ming-yuan, Li Hui, Chen Jia-tong 2010, 14 (14):  2533-2538.  doi: 10.3969/j.issn.1673-8225.2010.14.013 Abstract ( 115 )   PDF (1874KB) ( 349 )   Save BACKGROUND: Schwann cells can secrete various neurotrophic factors, and promote functional recovery of injured spinal cord. However, xenogenic Schwann cells transplantation may induce autoimmune response. Moreover, local transplantation results in secondary injury. Vein transplantation may reached injury site passing the blood spinal cord barrier, but the treatment concentration is not effective. OBJECTIVE: To investigate the therapeutic effects of transplantation of autologous activated Schwann Cells (AASCs) via subarachnoid space on spinal cord injury (SCI) in rats. METHODS: A total of 66 rats were used to establish SCI models, and the model rats were randomly divided into 3 groups. The unilateral saphenous nerves of rats were ligated directly for 1 week to activate Schwann cells, but inactivated and model control groups were not subjected to nerve ligation. 1 cm nerve was excised from distal end of each group, and Schwann cells were isolated and cultured by tissue mass method. The AASCs, autologous Schwann cells (ASCs) were injected with corresponding Hoechst33342-labeled SCs suspension, but the model control group was injected with DMEM injection. The basso beattie bresnahan (BBB) score and footprint analysis, as well as HE and GFAP immunohistochemistry staining were performed to evaluate functional recovery of rat hind limbs. RESULTS AND CONCLUSION: On 4 weeks after injury, BBB scores of AASCs were significantly superior to the other groups (P < 0.05). Two weeks after transplantation, some SCs migrated to injured spinal cord. Compared with ASCs group, the center distance of forward and hind feet and extorsion angle of the third toe of hind limb were significantly reduced in the AASCs group at 5 weeks (P < 0.05), the glial scar area was significantly decreased at 13 weeks (P < 0.05), and the cavity area of injured region was significantly diminished (P < 0.05). Results show that AASCs transplantation via subarachnoid space promoted functional recovery after SCI in rats. Related Articles | Metrics

Select

Stereotaxic intracerebral transplantation of neural stem cells with Nogo-66 receptor gene silencing for treating traumatic brain injury in rats  Wang Dong, Zhang Jian-jun, Ma Jing-jian 2010, 14 (14):  2539-2544.  doi: 10.3969/j.issn.1673-8225.2010.14.014 Abstract ( 106 )   PDF (1574KB) ( 440 )   Save BACKGROUND: Neural stem cells (NSCs) have the potential of self-proliferation and multiple directional differentiation, and can differentiate into various cells in the neural system under a certain condition. Therefore, NSCs have good prospect in repair of nerve injury. However, RNA interference avoids the abuse of permanent gene silencing, and is hopeful to combine with NSC transplantation for treating craniocerebral injury. OBJECTIVE: To determine whether the Nogo-66 receptor (NgR) gene silencing in NSCs can enhance curative effects of stereotaxic intracerebral transplantation of NSCs on traumatic brain injury (TBI) in rats. METHODS: A total of 60 male Wistar rats following TBI establishment were randomly assigned to 3 groups (n = 20). In the experimental group, NgR gene silencing NSC suspension (6 μL) was injected into rat brain tissue following 24 hours of model induction. In the control group, an equal volume of NSC suspension was infused by the same method. In the blank group, an equal volume of medium without stem cells was infused by the same method. At 24 hours, 3 days, 1 and 2 weeks following injury, neurological deficits were scored. Two weeks later, animals were sacrificed and subjected to immunohistochemistry and hematoxylin-eosin staining. RESULTS AND CONCLUSION: Following transfection of small interfering RNA, compared with control group, NgR gene protein expression was significantly reduced in the experimental group. At 1 and 2 weeks following transplantation, neurological deficit score was significantly less in animals undergoing NSC transplantation in the experimental group compared with the control group (P < 0.05). Moreover, neuron number in the brain tissue sections of experimental group was significantly more than in the control group (P < 0.01). At 2 weeks following injury, hematoxylin-eosin staining showed that brain tissue breakage at damaged site as scar connection, remarkable porosis in the blank group; typical morphological changes as neural cells at the transplanted site in the control group; typical morphological changes as neural cells without cavity in the experimental group. Immunohistochemistry showed (37.92±16.02) BrdU-labeled positive cells/high-power field in the blank group, (89.68±15.34) cells/high-power field in the control group, and (102.67±13.52) cells/high-power field in the experimental group. Significant differences were detected between groups (P < 0.01). The above-mentioned results indicated that the NSCs of NgR gene silencing transplanted into the injured cerebral tissues can significantly improve the neurological function in rats with TBI. Related Articles | Metrics

Select

Evaluation of autologous peripheral blood stem cell transplantation for diabetes combined with lower limb artery obturation using color Doppler ultrasound: Identical to results from angiography? Zhou Fang-ping, Chang Hong-xian, Yao Xiu-song, Chen Jing, Zhang Hui, Jiang Kun, Sun Hong, Liu Jian, Sui Yang 2010, 14 (14):  2545-2548.  doi: 10.3969/j.issn.1673-8225.2010.14.015 Abstract ( 142 )   PDF (571KB) ( 374 )   Save BACKGROUND: Traditional therapeutic methods of diabetes combined with lower limb artery obturation showed poor outcomes such as drug or intervention. Stem cell transplantation is a new method to treat this disease in recent years, and has obtained good outcomes in the clinic. Evaluating the therapeutic outcomes of stem cell transplantation has great damage and cost much. Thus, color Doppler ultrasound is used to examine non-invasion in the artery of the lower extremity, which shows important clinical value. OBJECTIVE: To evaluate improvement of lower extremity artery lesion following autologous peripheral blood stem cell transplantation using color Doppler ultrasound, and to compare with results of angiography. METHODS: A total of 41 patients diagnosed as having diabetes combined with lower limb artery obturation were included. Following administration of recombinant human granulocyte-colony stimulating factor (rhG-CSF) (150 µg/d) for 5 days, peripheral blood mononuclear cells (PBMNCs) were mobilized and harvested by using a separator at day 5 to prepare stem cell suspension, and which were injected intramuscularly in the diseased extremities (3×109 per limb). The clinical and laboratory findings were monitored every week for 3 months to 1 year following stem cell transplantation. RESULTS AND CONCLUSION: After stem cell transplantation, severe pain lameness, local cool-feeling and ulcer were significantly improved, and ankle-brachial index was increased obviously (P < 0.01). Color Doppler ultrasonography measurement showed that peak systolic velocity in diseased extremities was significantly increased post-transplantation (P < 0.01). Following transplantation, blood flow of the lower limb arterial canal was improved to different extents; collateral vessels were increased surrounding obstructed artery and in muscles. Spiral CT for new collateral vessel formation were showed increased. No related complication or adverse effects were observed in 41 patients during observation. Results indicated that color Doppler ultrasound can exactly rapidly examine and assess blood transport improvement in the lower limbs following stem cell transplantation, which is identical to results from angiography. It also provides reliable evidences for the diagnosis on the transplantation of stem cells for the treatment of diabetes combined with lower limb artery obturation. Related Articles | Metrics

Select

Stereotaxic transplantation of bone marrow mesenchymal stem cells in treatment of spinal cord injury in rats Guan Ya-lin, Kong Fan-ming, Wang Shi-min, Wu Sheng, Wang Wan-jun, Tang Fan, Zhang Wen-zhi 2010, 14 (14):  2549-2555.  doi: 10.3969/j.issn.1673-8225.2010.14.016 Abstract ( 65 )   PDF (708KB) ( 444 )   Save BACKGROUND: The key of stem cells for treating nervous tissue injury is the transplantation of stem cells that have regeneration capacity. The structure and function of central nervous system were re-established by multiple action mechanisms. OBJECTIVE: To explore the effects and mechanisms of bone marrow mesenchymal stem cells (BMSCs) locally transplanted into rats with spinal cord injury on neurological recovery. METHODS: BMSCs were separated with density gradient centrifugation and cell attachment. 10 mg/L BrdU was used for labeling before cell transplantation. Adult female Wistar rats were used to establish spinal cord injury models using an aneurysm clip, and they were then randomly divided into control group, saline group and transplantation group. In the transplantation group, BMSCs were transplanted into the damaged spinal cord by stereotaxis at day 7 following damage. In the saline group, an equal volume of saline was utilized. In the control group, the rats were left intact. Basso, Beattie and Bresnahan (BBB) locomotor rating scale was used before and at 7, 14, 30, 60 and 90 days following damage. Rats were sacrificed at day 90. BrdU-positive cells, Brdu+neuron specific enolase, Brdu+glial fibrillary acidic protein (GFAP), Brdu+basic fibroblast growth factor (bFGF), and Brdu+brain-derived nerve growth factor (BDNF) immunohistochemistry double-staining cells and simple staining positive cells were observed. RESULTS AND CONCLUSION: The recovery of BBB function score was better in the transplantation group than in the control group (P < 0.05). The recovery speed of BBB function score was slower in the saline group than in the control group at 30 days following damage (P < 0.05). No significant difference was determined at day 90 compared with the control group (P > 0.05). BrdU-positive cells and double-staining cells of immunohistochemistry could be found at the center of damage site and 1 cm from caudal end to damaged site in rats of the transplantation group. The number of NSE, GFAP, bFGF and BDNF simple staining cells was significantly greater in the transplantation group than in the control and saline groups (P < 0.05). Results indicated that BMSC transplantation can improve the recovery of nervous function of rats with spinal cord injury. Its mechanism may be correlated with the differentiation of transplanted cells into neuron-like and glial cell-like cells, secretion or promoting secretion of neurotrophic factors in host. Related Articles | Metrics

Select

Local injection of bone marrow mesenchymal stem cells for spinal cord injury in rats: Is motor function improved?  Guo Mian, Zheng Yong-ri, Li Qing-song, Wang Jian-jiao, Sun Jia-hang, Ge Yun-long, Zhao Yan 2010, 14 (14):  2556-2559.  doi: 10.3969/j.issn.1673-8225.2010.14.017 Abstract ( 100 )   PDF (1565KB) ( 342 )   Save BACKGROUND: Present studies mainly focused on in vitro culture of bone marrow mesenchymal stem cells (BMSCs) and cell transplantation for treating intracalvarium diseases. However, the understanding of survival, differentiation, migration and structure of transplanted cells in the damaged spinal cord is limited. OBJECTIVE: To explore effects of local BMSC transplantation in repair of spinal cord damage and feasibility of replacement therapy of BMSCs. METHODS: Adult healthy female Sprague-Dawley rats were randomly assigned to cell transplantation and control groups. Rat models of spinal cord transection damage were established. Rat BMSC suspension or calcium and magnesium phosphate buffer were transplanted immediately after injury to the damage zone. At 1 day, 1, 2, 3, 4 and 8 weeks before and after transplantation, BBB score motor function was observed in rats, and at 1 week after transplantation, immunohistochemical staining was utilized to observe BrdU-labeled BMSC survival in the spinal cord damaged site. At 4 weeks after transplantation, the general observation and histological detection were observed. RESULTS AND CONCLUSION: At 1-8 weeks after transplantation, BBB scores were higher in the cell transplantation group than in the control group. At 1 week following surgery, immunohistochemical staining showed that BrdU-positive cells were detected in the distal end of rat spinal cord in the cell transplantation group. At 4 weeks following surgery, nerve fibers were found in the damaged spinal cord. These verified that BMSCs were transplanted into rat damaged spinal cord immediately following damage, and the transplanted cells could survive. Living BMSCs can differentiate into neurons, and formed neuron pathway in the local region of damage, which will promote the recovery of conduction function of spinal nerve fibers, and contribute to the recovery of rat hindlimb motor function following high-level spinal cord injury. Related Articles | Metrics

Select

Bone marrow-derived mesenchymal stem cells injection for liver failure in New Zealand white rabbits Ling Xiao-hua, Hu Cheng-yi, Hong Yu, Yu Xin, Mi Li-na, Yang Xiao-ming 2010, 14 (14):  2560-2563.  doi: 10.3969/j.issn.1673-8225.2010.14.018 Abstract ( 93 )   PDF (1161KB) ( 403 )   Save BACKGROUND: Hepatocyte transplantation as an effective method for liver failure has been confirmed by animal models and clinical application. However, limited source and poor proliferation of hepatocyte graft limit its development. Studies have shown that bone marrow mesenchymal stem cells (MSCs) have potentials to differentiate into hepatocyte and bile epithelial cells, with strong proliferation. OBJECTIVE: To explore the therapeutic effect of bone marrow MSCs transplantation on liver failure of New Zealand white rabbits. METHODS: Adult male New Zealand rabbits were treated with D-galactosamine, and 3 mL hepatocyte suspension (1×109 /L) was injected into the liver of transplantation group, but the control group was injected with the same volume of culture solution with no bone marrow MSCs. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity was detected 48, 72 hours, 1, 4 weeks following transplantation, and pathological detection was performed at 4 weeks. RESULTS AND CONCLUSION: The liver functional index following transplantation of bone marrow-derived MSCs transplantation was significantly decreased, and ALT and AST activity at 4 weeks was significantly less than the control group (P < 0.05). At 4, the transplantation group displayed disorderly hepatic cord, hepatocyte swollen and degeneration, necrosis, accompanied by bleeding and inflammatory cell infiltration. In addition, the hepatic lobule structure was detectable, and regenerative hepatocyte increased among necrotic hepatocyte; small cells with large ratio of nucleus and cytoplasm at header, central vein and surrouding necrosis focus extended to the liver tissues. Related Articles | Metrics

Select

Influence of astragalus injection on in vitro cultured olfactory ensheathing cell proliferation and neurotrophic factor secretion Wang Pei-min, Li Jian-feng, Zhang Nong-shan, Zhang Xu 2010, 14 (14):  2564-2567.  doi: 10.3969/j.issn.1673-8225.2010.14.019 Abstract ( 73 )   PDF (1320KB) ( 363 )   Save BACKGROUND: Astragalus or olfactory ensheathing cell (OECs) transplantation alone can promote survival of various neurons and functional improvement of injured spinal cord. The clinical effect of astragalus in combination with OECs transplantation remains unclear. OBJECTIVE: To observe the effect of astragalus on the proliferation of OECs and the secretion of neurotrophic factors.  METHODS: OECs of SD rats, 24 hours old, were isolated and cultured, and identified by immunohistochemistry. OECs at culture day 8 were seeded in polylysine-coated 96-well plate and incubated in 5% CO2 at 37 ℃ for 24 hours, followed by   2 mg/L, 20 mg/L, 200 mg/L, 2 g/L, 20 g/L astragalus injection for 24 hours. MTT and flow cytometry were used to assess the effects of astragalus in promoting OECs proliferation; enzyme linked immunoassay (ELISA) was used to measure the content of neurotrophic factors. Serum culture medium was served as negative control.  RESULTS AND METHODS: Compared with the negative control group, 2 and 20 mg/L astragalus injection significantly promoted OECs proliferation (P < 0.05, P < 0.01). The result of flow cytometry showed that astragalus could promote OECs in G1 stage and reduce the percentage of cells in S and G2 stages, but the difference was not significant (P > 0.05). However, astragalus injection at each mass concentration significantly reduced the number of OECs (P < 0.05). Moreover, 2 mg/L astragalus injection significantly promoted in vitro neurotrophic factor levels in OECs. Results show that astragalus injection in combination with OECs can promote recovery of spinal cord injury. Related Articles | Metrics

Select

Stem cell factor promotes the proliferation and osteogenic differentiation of dental pulp stem cells Xin Hong, Chen Run-liang, Pei Ying-bo 2010, 14 (14):  2568-2571.  doi: 10.3969/j.issn.1673-8225.2010.14.020 Abstract ( 119 )   PDF (1116KB) ( 404 )   Save BACKGROUND: Stem cell factor (SCF) serves as a factor having a promotion effect on proliferation and differentiation of stem cells. Whether SCF affects biological features of dental pulp stem cells (DPSCs), and plays an important role in tooth regeneration remain unclear. OBJECTIVE: To observe SCF effects on the proliferation and osteogenic differentiation of human DPSCs. METHODS: Pulp tissues were removed from healthy human teeth extracted for orthodontic purposes. The pulp tissues were digested by collagenase and dispase. The sample was cultivated in culture flasks with medium contained 1, 10 μmol/L SCF; the normal culture medium served as control. The cell proliferation ability was detected by MTT. The osteogenic gene bone sialoprotein and osteocalcin mRNA expression were examined by real-time PCR. Alkaline phosphatase activity was detected by alkaline phosphatase kit. RESULTS AND CONCLUSION: SCF could enhance the proliferation ability, upregulate bone sialoprotein and osteocalcin mRNA expression, and enhance the alkaline phosphatase activity, with the tendency of increased promotion with increased concentration. Results indicated that SCF could enhance the proliferation and osteogenic differentiation ability of human DPSCs. Related Articles | Metrics

Select

Nerve growth factor combined with neural stem cell transplantation for treating spinal cord injury in rats  Fan Guang-ming, Zhang Wen-bin, Zhang Sai, Wang Li-jun 2010, 14 (14):  2572-2578.  doi: 10.3969/j.issn.1673-8225.2010.14.021 Abstract ( 101 )   PDF (789KB) ( 511 )   Save BACKGROUND: The effects of simple neural stem cell (NSC) transplantation on repair of damaged spinal cord are not ideal. Previous studies have shown that nerve growth factor (NGF) has both effects of neuron nutrition and process growth promotion, can effectively contribute to the recovery of neurofunction following spinal cord injury (SCI). OBJECTIVE: To investigate the effect of neural stem cell transplantation combined with NGF application on the recovery of motor function of rats with SCI. METHODS: A total of 42 Sprague Dawley rats were used to establish SPI models, and then divided randomly into three groups: medium, simple NSCs or NSCs + NGF. At 1 week following injury, medium, simple NSCs or NSCs + NGF were separately injected into damaged sites. At 1, 2, 4, 6, 8 weeks post-injury, all animals were evaluated on the hind limb behavior with Basso, Beattie and Bresnahan (BBB) locomotor rating scale and inclined plane test. At 4 weeks post-transplantation, histopathology hematoxylin-eosin staining and BrdU immunohistochemistry were performed. At 8 weeks post-transplantation, horseradish peroxidase (HRP) nerve trace and somatosensory evoked potential testing were performed to observe the recovery of nerve electrophysiology. RESULTS AND CONCLUSION: At 4 weeks following injury, motor function of the rat hindlimb was significantly improved in the simple NSCs group and NSCs + NGF group. The recovery was rapid in the NSCs + NGF group than the simple NSCs group (P < 0.05). The recovery was slight in the medium group. Pathological sections demonstrated that neurite was not found in the medium group. A few neurites were seen in the simple NSCs group, and many neurites were observed in the NSCs + NGF group. Numbers of BrdU-positive cells and HRP-positive nerve fibers: NSCs + NGF group > simple NSCs group > medium group, there was significant difference among groups (P < 0.01). Latent period and amplitude of somatosensory evoked potentials in the NSCs + NGF group were superior to the simple NSCs group (P < 0.05), and significantly better than the medium group (P < 0.01). Results suggested that NSC transplantation can promote the recovery of hindlimb function, and combined with NGF presents synergic effect. Related Articles | Metrics

Select

Effect of basic fibroblast growth factor on the formation of blast-colony-forming cell derived from embryonic stem cells Li Na, Shi Zeng-li, Wang Yue-si 2010, 14 (14):  2579-2582.  doi: 10.3969/j.issn.1673-8225.2010.14.022 Abstract ( 138 )   PDF (638KB) ( 395 )   Save BACKGROUND: Some studies show that basic fibroblast growth factor (bFGF) strongly expresses during the process of embryonic stem cells differentiation into hematopoietic stem cell, yolk sac blooding, and fetal liver hematopoiesis. OBJECTIVE: To study the regulation of bFGF on the blast-colony-forming cell (BL-CFC) by adding bFGF in the medium of embryoid body generation. METHODS: The third to fifth generations of the primary mouse embryonic fibroblasts were recovered, and then incubated with the DMEM medium containing mitomycin C for 2.5 hours in order to lose the proliferative capacity. Then cells were suspended into single cell by trypsinization and inoculated in the gelatin-coated bottle at the density of 10×104/cm2. After culturing for 24 hours, mouse embryonic stem cells (mESC) of D3 were recovered and placed on the feeder layer cells. According to the composition of medium in embryoid body generation, mESCs were divided into two groups: group A: standard medium + VEGF + SCF; group B: standard medium + VEGF + SCF + bFGF. Each group was cultured for 3 days and 6 days respectively, and the cloning number of BL-CFC was quantified, as well as Flk-1+ expression was observed by immunofluorescence staining. Positive number and average absorbance were analyzed using IMAGE-PRO PLUS imaging analysis system. RESULTS AND CONCLUSION: Adding bFGF in the course of embryoid body growth could significantly increase the number of BL-CFC (P < 0.01), and the positive results of Flk-1 and the average absorbance were also increased significantly (P < 0.01). bFGF effectively promoted embryoid body amplification and proliferation of BL-CFC. Related Articles | Metrics

Select

Neuroprotective effect of glial cell line-derived neurotrophic factor gene-modified bone marrow stromal cells transplantation on intracerebral hemorrhage in rats  Deng Li, Yang Chao-xian, Tu Jiang-yi, Gao Xiao-qing, Guo Kan, Yuan Qiong-lan 2010, 14 (14):  2583-2587.  doi: 10.3969/j.issn.1673-8225.2010.14.023 Abstract ( 150 )   PDF (670KB) ( 433 )   Save BACKGROUND: Previous studies have demonstrated that cell transplantation has neuroprotective effect on intracerebral hemorrhage, and some researches have indicated that transplantation of bone marrow stromal cells (BMSCs) can promote neural function recovery after cerebral infarction. OBJECTIVE: To explore whether transplantation of BMSCs-modified by glial cell line-derived neurotrophic factor gene (GDNF) gene provides a better therapeutic effect than native BMSCs after stroke. METHODS: Totally 36 SD rats were induced intracerebral hemorrhage models by injecting autologous arterial blood, and then divided into 3 groups (n = 6), each group was assigned into 2 sub-groups. Rabbits in each group were stereotaxically grafted with 20 μL GDNF/BMSCs, BMSCs or saline respectively. The rats were executed at 1 and 2 weeks after operation, and immunohistochemistry was used to observe the expressions of synaptophysin (Syn) and growth associated protein-43 (GAP-43) in the margin of the hemorrhagic focus. RESULTS AND CONCLUSION: Compared with the BMSCs and control groups, both Syn-immunoreactive and GAP-43-immunoreactive products were significantly increased in the GDNF/BMSCs group (P < 0.05). Present results demonstrate that transplantation of GDNF gene-modified BMSCs provides better neuroprotection than native BMSCs delivery for stroke. Related Articles | Metrics

Select

Ad5-enhanced green fluorescent protein versus rAAV2-enhanced green fluorescent protein transfecting adipose-derived mesenchymal stem cells   Yuan Xiao-hong, An Rong-ze, Wang Zhao-jie, Jia E-nuo, Qi Xin-wen, Chen Jin-ping, Yang Jin, Liu Fan-fan 2010, 14 (14):  2588-2590.  doi: 10.3969/j.issn.1673-8225.2010.14.024 Abstract ( 175 )   PDF (1010KB) ( 296 )   Save BACKGROUND: There are reports concerning differentiation of adipose-derived mesenchymal stem cells (ADSCs) into chondrocytes using gene transfection technique. However, the transfection of adenovirus and adeno-associated virus into ADSCs is various. It is controversial whether adeno-associated virus (AAV) can transfect ADSCs. OBJECTIVE: To observe the enhanced green fluorescent protein (EGFP) expression following Ad5-EGFP and rAAV2-EGFP transfection into ADSCs, and investigate the cell proliferation ability following transfection. METHODS: ADSCs were isolated from the adipose tissue, which was from 6-month-old New Zealand albino rabbit back and neck by mechanical digestion and enzyme digestion, then ADSCs were cultured and amplified in vitro. ADSCs were infected with Ad5-EGFP and rAAV2-EGFP, and the EGFP expression was observed. A total of 2 μL sodium butyrate (1 mol/L) was added into the medium after rAAV2-EGFP transfection. MTT assay was used to detect the gene transfection effects on reproductive activity of ADSCs. RESULTS AND CONCLUSION: ADSCs isolated and cultured in vitro were flat, long-spindle and amplified stably. The cell morphology was uniform. Many green fluorescent cells were observed in Ad5-EGFP and rAAV2-EGFP groups. Transfection efficiencies were about 88% and 83%. Adenovirus and adeno-associated virus vector can be transfected with ADSCs, and transfection efficiency is high. Adeno-associated virus needs sodium butyrate to increase its level of gene expression. Related Articles | Metrics

Select

In vitro culture and surface marker variations of umbilical cord blood mesenchymal stem cells Liu Su-fang, Duan Dong-xiao, Han Xue-fei, Yan Wen-hai, Xing Ying 2010, 14 (14):  2591-2595.  doi: 10.3969/j.issn.1673-8225.2010.14.025 Abstract ( 93 )   PDF (1059KB) ( 329 )   Save BACKGROUND: Currently, there is not a standard method for in vitro culture and large scale amplification of umbilical cord blood mesenchymal stem cells (UCB-MSCs). OBJECTIVE: To investigate the isolation, purification and culture of UCB-MSCs in vitro, and to detect its surface marker variation. METHODS: The monocytes were harvested from UCB using 1.077 g/cm3 lymphocytes separating solution and density gradient centrifugation, followed by incubation in an incubator containing 5%CO2 at 37 ℃. The cell morphological changes were observed at different time points and the expression of surface marker was detected using flow cytometry.  RESULTS AND CONCLUSION: The monocytes isolated from the UCB grew initially into numerous hematopoietic cell clones, most of which were granulocyte/macrophage colony-forming units and burst forming unit-erithroid, increasing by (37.1±2.3) and (10.4±1.7), respectively. Switzerland staining showed most of them were granulocyte clones (80.1±85.2)%, next was erythroid clones (14.2±1.8)%. At 7 days after culture, some shuttle fibroblast-like cells and flat osteogenic-like cell spread the whole plastic well. At 14 days after culture, flow cytometry showed CD38+ cells accounted for 1.64%, and CD34+ /CD38+ cells accounted for 1.71%, and CD34+ /CD38- were 0.55%. PI+ and Annexin-V+ cells accounted for 0.05% and 0.18% respectively. At 21 days after culture, CD38  + , CD34+ /CD38+ and CD34+ /CD38- cells were 74.32%, 1.61%, and 0.24%. The results reveled that UCB-MSCs can be isolated and cultured in vitro. Related Articles | Metrics

Select

Isolation, purification and cultivation of rat muscle-derived stem cells  Ye Jin, Jin Feng-shuo, Chen Jin, Wang Peng, Liang Pei-he, Nie Zhi-lin, Li Qian-sheng 2010, 14 (14):  2596-2600.  doi: 10.3969/j.issn.1673-8225.2010.14.026 Abstract ( 116 )   PDF (1204KB) ( 359 )   Save BACKGROUND: In vitro screening and amplification are important links to harvest muscle-derived stem cells that are satisfactory to clinical requirement. OBJECTIVE: To probe into the method of isolation, culture and purification of skeletal muscle-derived stem cells from adult rats in vitro. METHODS: The skeletal muscle was obtained sterilely following adult Sprague Dawley rats were anesthetized. Muscle-derived stem cells were harvested using enzyme digestion with XI collagenase, Dispase and trypsogen, and then purified by Percoll density gradient centrifugation and differential adhesion method. Growth curves were recorded and MTT colorimetric technique was used to describe the effects of various kinds of inoculum density on cell growth. Cells were identified by immunocytochemistry. RESULTS AND CONCLUSION: Primary muscle-derived stem cells were less in volume, lower adherence and well refraction, appearing as globular or fusiform or spindle and slowly multiplication. Following subculture, complete medium containing 20% serum was added. Cell number was greatest when cell density was 1×109/L, which was the optimal density. Cells at passages 1-4 grew well. Cells showed desmin(+), CD34(+), CD45(-) and Sca-1(+) by immunocytochemistry. Results verified that high-purity muscle-derived stem cells can be obtained in vitro and amplified successfully following primary culture. Related Articles | Metrics

Select

Isolation, culture and multi-directional induced differentiation of rabbit adipose derived stromal stem cells   Yang Hao, Wu Di, Li Shi-he, Zhu Xiao-song 2010, 14 (14):  2601-2606.  doi: 10.3969/j.issn.1673-8225.2010.14.027 Abstract ( 157 )   PDF (1204KB) ( 414 )   Save BACKGROUND: Subcutaneous fat of human body is a rich reservoir of adipose derived stromal stem cells (ADSCs). ADSCs can proliferate rapidly when being cultured in vitro, and has the capacity of multi-directional differentiation. ADSCs attracted much attention in research of tissue engineered seed cells. OBJECTIVE: To isolate and culture stromal vascular fraction (SVF) cells from rabbit subcutaneous fat in vitro, and to testify whether it has multiple differentiation capacity.  METHODS: SVF cells were isolated in vitro from rabbits, and cultured under standard condition. Cellular surface antigens CD44, CD45 and CD29 of passage 3 SVF cells were examined using flow cytometry. Passage 3 SVF cells were induced to differentiate into osteoblasts, chondrocytes, and lipocytes. Oil red staining was used to examine lipocyte induction. Alkaline phosphatase (ALP) staining, alizarin red staining and von Kossa staining were used to examine osteoblast induction. Type Ⅱ collagen immunohistochemical staining and type Ⅱ collagen mRNA RT-PCR were used to examine chondrocyte differentiation. RESULTS AND CONCLUSION: Primary SVF cells were multi-angular or short spindle-shaped. Passage 3 SVF cells were long spindle-shaped. Flow cytometry showed CD44+ , CD29+ , CD45- . Oil red staining exhibited positive reaction in lipocyte induction group. ALP staining, alizarin red staining and Von kossa staining demonstrated positive reactions in osteoblast induction group. Type Ⅱ collagen immunohistochemical staining and alcian blue staining have suggested positive reactions at 14 days of chondrogenic induction group. RT-PCR of type Ⅱ collagen mRNA test showed that the product band had strong signal at 14 days of chondrogenic induction group compared with that before induction. Above mentioned results have indicated that SVF cells isolated from rabbit subcutaneous fat have identical surface makers of stem cells, and have the ability to differentiate into lipocytes, osteoblasts and chondrocytes in vitro by induction, and it could be concluded that the SVF cells were ADSCs. Related Articles | Metrics

Select

Effects of autologous platelet-rich plasma on function and activity of human bone marrow-derived endothelial progenitor cells   Zhao Zhong-hai, Li Hong-qiu, A Liang, Chen Bin, Wang Shao-bo 2010, 14 (14):  2607-2611.  doi: 10.3969/j.issn.1673-8225.2010.14.028 Abstract ( 130 )   PDF (1159KB) ( 432 )   Save BACKGROUND: Platelet-rich plasma is the plasma containing rich platelet extracted by special method. Compared with common serum, platelet-rich plasma contains richer cytokines, such as platelet derived growth factor, transforming growth factor beta and vascular endothelial growth factor. OBJECTIVE: To observe effects of platelet-rich plasma on biological characteristics of human endothelial progenitor cells (EPCs), and to discuss the repair function of platelet-rich plasma on bone defects. METHODS: Platelet-rich plasma was collected from autologous peripheral vein blood of 16 healthy volunteers. EPCs were harvested from bone marrow blood by density gradient centrifugation. EPCs following 8 days of culture were randomly assigned to control group, platelet-rich plasma group, fetal bovine serum group and serum-free control group. In the fetal bovine serum group, cells were incubated in high-glucose DMEM containing 10% fetal bovine serum, 100 U/mL penicillin and 100 U/mL streptomycin. In the serum-free control group, cells were incubated in high-glucose DMEM containing 100 U/mL penicillin and 100 U/mL streptomycin. EPC growth was observed under a phase contrast microscope and a laser confocal microscope. AC133- and vWF- positive cells (double staining) were differentiated EPCs. EPC proliferation, migration and tubule formation capacity were respectively measured by the MTT assay, Transwell chamber assay and Matrigel tubule assay at 6, 12, 48 hours following culture. RESULTS AND CONCLUSION:  ① Cells from each group began to adhere to the wall, and changed from round to spindle, polygonal and irregular shapes at 12-24 hours following incubation. ② A490 value in EPCs was significantly greater following 6 hours of treatment with platelet-rich plasma compared with the control group (P < 0.05), and the promotion effect became stronger at 12 hours (P < 0.01). At 0-48 hours, with prolonged time, the promotion effect of platelet-rich plasma on EPC proliferation enhanced. ③ Platelet-rich plasma could elevate EPC migration, and the effect became significantly enhanced at 6 hours (P < 0.05), peaked at 12 hours (P < 0.01), decreased at 48 hours, and still significantly greater than the control group (P < 0.01). ④ Platelet-rich plasma obviously enhanced tubule formation capacity in EPCs in vitro, especially at 48 hours, and the tubule-like structure is more complicated compared with the control group. Related Articles | Metrics

Select

Immunohistochemical analysis of the expression of Hes1 in adult mouse brain Huan Lin-chun, Yang Xin-yu, Zhao Wang-miao, Zhao Yan, Zhao Jie, Zhang Qi, Yang Shu-yuan 2010, 14 (14):  2612-2616.  doi: 10.3969/j.issn.1673-8225.2010.14.029 Abstract ( 126 )   PDF (1393KB) ( 544 )   Save BACKGROUND: In recent years, it has been shown in studies that regulation of the expression of Hes1 is very important to maintain neural stem cells and regulate its differentiation during the development of embryonic nervous system. Moreover, upregulation of Hes1 expression is required for quiescent adult fibroblasts resuming proliferation. This indicates that there is a close relationship between Hes1 and some potential stem cells in adult. Therefore, the study of Hes1 expression in neural stem cells and its relationship with adult neurogenesis is put on the agenda. However, the expression of Hes1 in adult brain is still not clear. OBJECTIVE: To observe the expression of Hes1 in adult mouse brain and the cell types of Hes1 positive cells. METHODS: Totally 12 adult male C57BL/6 mice were randomly evenly divided into two groups by using the random number table. For the first group, the mouse brain was removed directly and immunohistochemical staining was used to detect the expression of Hes1 in the adult mouse brain. For the second group, Brdu was injected intraperitoneally to every mouse with a dosage of 200 mg/kg, once a day, for 3 days. The mouse brain was removed at 4 days and then double staining was carried out to observe and analysis the cell types of Hes1 positive cells in hippocampus. RESULTS AND CONCLUSION: Hes1 was expressed in all the observed anatomical site where neural cells exist, Brdu positive cells nearly all expressed Hes1, NeuN positive cells all expressed Hes1, whereas GFAP positive cells did not express Hes1 completely. Hes1 was expressed in all neurons and neural stem cells, and glial cells did not express Hes1. Related Articles | Metrics

Select

Primary culture of spiral ganglion cells from cochlea of neonatal C57 mice  Wang Ting-kuo, Sun Hong 2010, 14 (14):  2617-2620.  doi: 10.3969/j.issn.1673-8225.2010.14.030 Abstract ( 227 )   PDF (1218KB) ( 473 )   Save BACKGROUND: At present, there are differences in reporting culture condition of cochleal spiral ganglion cells (SGCs) in circle of primary cell culture. Individual method has poor repeatability, and is not beneficial for practical application. OBJECTIVE: To primary culture and evaluate SGCs of neonatal C57 mice. METHODS: Tissues from cochlear modiolus were acquired from neonatal C57 mice by microanatomy. SGCs from cochlear modiolus were cultured by trypsin digestion, differential velocity adherent technique combined with chemicals. Cell growth was observed by inverted phase contrast microscope and hematoxylin and eosin staining. Immunocytochemistry was employed to classify cell types.  RESULTS AND CONCLUSION: After purification, the cell body of SGCs from cochlear modiolus was ellipse or triangle, with slender processes in cytoplasm. Nuclei were positive for Nuen (stained brown). Cytoplasm and axon were positive for β3-Tubulin (stained brown). These indicated that mouse SGCs were successfully cultured. Related Articles | Metrics

Select

Position and application of proteomics in induced differentiation of bone mesenchymal stem cells Hu Feng, Zhao Jin-min 2010, 14 (14):  2623-2626.  doi: 10.3969/j.issn.1673-8225.2010.14.032 Abstract ( 68 )   PDF (553KB) ( 343 )   Save BACKGROUND: The essence of cell differentiation is a selectively intra-cellular gene expression, which results in specific proteinic synthesis and causes changes in biochemistry, structure and function. Thus, original proteomics and a single protein analysis can not meet the requirement in study. Proteomics technology provides a powerful tool due to the large scale, systemical study of protein transformation and interaction, which can be used for exploring molecular mechanism of bone mesenchymal stem cells (BMSCs) during directional differentiation. OBJECTIVE: To introduce proteomics, to summarize the research of proteomics in directional differentiation of BMSCs, and to forecast the development of proteomics research methods. METHODS: To search articles highly related with BMSCs, cell differentiation, and proteomics published on CNKI (www.cnki.net/index.htm), Sciencedirect (http://www.Sciencedirect.com), I.S.I (http://www.isiwebofknowledge.com) were searched, and the key achievements were included in the analysis. RESULTS AND CONCLUSION: A total of 29 documents were reviewed, and the experiences in the application of proteomics technology in the directional differentiation of BMSCs were summarized. With the innovation and development in methodology and technology, proteomics will become a powerful tool for us to study the potential mechanisms of BMSCs directional differentiation. Related Articles | Metrics

Select

Role of bone morphogenetic protein 2 and DLX5 in the differentiation of neural stem cells from anterior subventricular zone into neurons Liu Sheng-hua, Zhou Zheng, Feng Bao-hai 2010, 14 (14):  2627-2630.  doi: 10.3969/j.issn.1673-8225.2010.14.033 Abstract ( 119 )   PDF (571KB) ( 437 )   Save BACKGROUND: Regulation of neural stem cells differentiation hampers its application, and the regulation mechanism remains poorly understood. Anterior subventricular zone (SVZa) is one of the rich zones of neural stem cells. The neural stem cells at SVZa can immigrate for a long distance with undifferentiated states and finally differentiate into interneurons at the olfactory bulbs. Bone morphogenetic protein-2 (BMP2) and DLX5 play a significant role in immigration and differentiation of neural stem cells at SVZa towards olfactory bulb. OBJECTIVE: To study the feature of construction and immigration of neural stem cells at SVZa, and to review the role of BMP2 and DLX5 in neural stem cells differentiation, in addition, to explore the regulation mechanisms of neural stem cells differentiation. METHODS: Literatures from PubMed database (http://www.ncbi.nlm.nih.gov/PubMed), CNKI database (www.cnki.net/index.htm) and WANFANG database (http://www.wanfangdata.com.cn) published between January 1997 and January 2009 were searched with the key words of “neural stem cells, SVZa, BMP2, and DLX5”. The repetitive and obsolete studies were excluded.  RESULTS AND CONCLUSION: A total of 121 literatures were selected and primarily collected, including 86 Chinese literatures and 35 English literatures. Finally, according to standardization, 28 literatures were further analyzed. Neural stem cells of SVZa have unique construction and can immigrate for a long distance. BMP2 and DLX5 play a significant role in neuronal immigration and differentiation of the NSCs of SVZa, but the detailed mechanism of BMP2 and DLX5 in SVZa is not clear. Related Articles | Metrics

Select

Application and progress of neural stem cells in spinal cord injury Chen Jia-li, Zhou Ji-hui, Lang Jin-bo 2010, 14 (14):  2631-2634.  doi: 10.3969/j.issn.1673-8225.2010.14.034 Abstract ( 83 )   PDF (562KB) ( 388 )   Save BACKGROUND: Neural stem cells (NSCs) are characterized by widely resources, convenient harvesting, easy culture and prone to import and express exogenous genes, which can be served as carriers of gene therapy for neural system disease. OBJECTIVE: To review the application of NSCs in spinal cord injury. METHODS: Databases of PubMed (http://www.ncbi.nlm.nih.gov/PubMed) and Wanfang (http://www.wanfangdata.com.cn) were searched by the correspondence author using key words of “neural stem cells, spinal cord injury, cellular transplantation” both in English and Chinese to retrieve papers concerning isolation, identification, differentiation of NSCs as well as its application in repairing spinal cord injury. A total of 82 documents were initial obtained by computer, after repetitive studies were excluded, 23 papers were included in the final analysis.  RESULTS AND CONCLUSION: The NSCs transplantation has been widely used in animal experiments. Currently, the applications of NSCs in repairing spinal cord injury are concentrated on the following aspects: Firstly, cellular replacement therapy, namely, direct transplanting NSCs or activating in vivo NSCs to differentiate into neurons and glial cells, and then integrating transplanted cells with the existed neural cellular structure to cure the disease. Secondly, NSCs were utilized as gene carriers, which carrying target gene to body and reach the aims of cellular replacement and gene therapy. Thirdly, autologous NSCs were induced differentiation for self neural repair via studying growth factors and cytokines. Related Articles | Metrics

Select

Research and application features of stem cells in dentistry Sha Ri-na, Wu Yan 2010, 14 (14):  2635-2637.  doi: 10.3969/j.issn.1673-8225.2010.14.035 Abstract ( 92 )   PDF (473KB) ( 447 )   Save BACKGROUND: The dental pulp stem cells, marrow mesenchymal stem cells and periodontal ligament stem cells belong to the adult stem cells from oral cavity, which own two features of the adult stem cells: the ability of renewing themselves strongly and the potential of splitting. Through the continuous study of many kinds of stem cells from oral cavity, it is very helpful for human to show the process of making the firm organizations of tooth and the inducing mechanism, in order to direct the related clinical work and the prevention work. OBJECTIVE: To summarize features and application perspective of various adult stem cells from oral cavity in the clinic. METHODS: The first author retrieved PubMed database (http://www.ncbi.nlm.nih.gov/PubMed) and China National Knowledge Infrastructure (http://www.cnki.net) for relevant literatures published from January 1999 to December 2009. The key words were “stem cell, dentistry”. A total of 80 literatures were screened, and studies on basic research and clinical application of stem cells in stomatology medical field were included. Old and duplicated studies were excluded and finally 19 literatures were included. RESULTS AND CONCLUSION: With development of study, adult stem cells in oral cavity will be gradually found to establish integrated stem cell database, which can provide cell source for tissue engineering study in the oral cavity. Related Articles | Metrics

Select

Clinical application of modified BU/CY pretreatment scheme to peripheral blood hematopoietic stem cell transplantation Chen Xiao-xia, Wang Zhi-ming, Luo Xian-sheng, Xu Dan-dan, Li Xing, Lei Mei-qing 2010, 14 (14):  2638-2642.  doi: 10.3969/j.issn.1673-8225.2010.14.036 Abstract ( 282 )   PDF (695KB) ( 469 )   Save BACKGROUND: In allogene hematopoietic stem cell transplantation, the choice of preconditioning scheme is an important link of the success of hematopoietic stem cell transplantation, and a major research direction of stem cell transplantation. The myeloablative pretreatment scheme has great toxicity, and pretreatment related death rate is high. Thus, it is necessary to explore an ideal pretreatment scheme to expect a decrease in side effects and relapse. OBJECTIVE: To observe the effect of modified Bu/CY pretreatment regimen for treating hematologic malignancies. METHODS: The 8 patients were selected at the Department of Hematology, Haikou Municipal People’s Hospital Affiliated to Xiangya School of Medicine, Central South University from November 2003 to March 2008, and received modified Bu/CY pretreatment: cytarabine 2.0-3.0 g/(m2•d)× 2 d, intravenous drip, for 24 consecutive hours; myleran 4 mg/(kg•d)× 3 d; cyclophosphamide 50 mg/(kg•d)× 2 d; methyl-cyclohexyl nitrosourea 25 mg/(m2•d)×1 d; antithymocyte globulin 25 mg/(kg•d)× 4 d. We have increased the arabinosylcytosin dose twice, and changed to a 24-hour infusion via intravenous drip, so that pretreatment strength was increased and promote lasting implantation of hematopoietic stem cells, based on the modified program. Graft-versus-host disease (GVHD) prevention: cyclosporin A and mycophenolate mofetil would be used advanced to minus 7 days (one day before stem cell transfusion is minus 1) based on the classic methotrexate regimen. The ABO blood group changes and DNA were tested in patients before and after transplantation. RESULTS AND CONCLUSION:  ①The detection of hematopoietic reconstitution after transplantation: All patients have received hematopoietic reconstitution, with no pretreatment-related death. The white blood cells reduced to 0 after -3 - +7 days of hematopoietic stem cell transplantation, and continues to (3-22) days, +10 - +21 days white blood cells > 1.0×109 /L, +11 - +51 days, platelets > 20×109 /L. ②Incidence of GVHD: of 8 patients, there were GVHD Ⅳ grade (intestinal) in 1 case, acute graft-versus-host disease grade Ⅰ-Ⅱ in 3 cases. Above-mentioned results indicated that the further modification of BU/CTX2 regimen may be an effective pretreatment program, with a few side effects, which is better than the classic total-body irradiation/CY regimen. What’s more, it is simple accurate, reliable role of anti-leukemia and will be a safe and effective method for treating hematologic malignancies. Related Articles | Metrics

Select

Surface marker changes during induced differentiation of bone marrow mesenchymal stem cells into neuronal-like cells Ma Lan, Li Ke 2010, 14 (14):  2643-2646.  doi: 10.3969/j.issn.1673-8225.2010.14.037 Abstract ( 99 )   PDF (305KB) ( 314 )   Save BACKGROUND: Adult central nervous system lacks the ability to regenerate, so it is of great significance to find a new source of neural stem cells. OBJECTIVE: To investigate the differences between basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) in inducing bone marrow mesenchymal stem cells (MSCs) to differentiate into neurons in vitro. METHODS: MSCs were isolated from normal human bone marrow using density gradient centrifugation and cell attachment method. MSCs were plated in 96-well culture plates at a density of 0.25×108/L and cultured with 200 μL DMEM/F12 for 0, 1, 2, 3, 4, 5, 6, and 7 days, with 20 μL MTT (5 mg/mL) in 5 wells at each time point. The supernatant was removed and 100 μL dimethyl sulphoxide was added to each well for 4 additional hours of incubation. In addition, some cells were exposed to bFGF and EGF. Growth curve was determined with MTT method. Telomerase activity were examined by TRAP(PCR)-ELISA. Additionally, the functional differences of the two cytokines were checked by RT-PCR. RESULTS AND CONCLUSION: RT-PCR revealed that nestin, glial fibrillary acidic protein (GFAP) and neurofilament subunit M (NF-M) mRNA were expressed in un-induced MSCs of passage 4. Nestin expression reduced at 7 days. The expression of micro-tubule-associated protein-2 (MAP2) mRNA was not detected until the induction, and increased thereafter. The expression of MAP2 mRNA was greater in bFGF+EGF and bFGF alone groups compared with EGF alone group, and the expression of GFAP in EGF alone group was greater than other groups. Results showed that MSCs can be cultivated, proliferated and differentiated into neural stem cells in vitro. The differentiated neural stem cells have the activity of proliferation, but not have the ability of infinite proliferation as tumor cells.   Related Articles | Metrics

Select

Primary culture of rat retinal and visual cortical neurons: Methodological characteristics Zhang Di, Wu Xiao-ying, Liu Shuang-zhen 2010, 14 (14):  2647-2651.  doi: 10.3969/j.issn.1673-8225.2010.14.038 Abstract ( 90 )   PDF (386KB) ( 468 )   Save BACKGROUND: Primary culture of neurons is an important way to study the structure and functions of the nervous system. It is also important to explore pathomechanism and medicine reaction of some ophthalmology diseases. OBJECTIVE: To explore an optimal way to the separation and culture of retinal and visual cortical neurons in new-born rats through comparative observation of different primary culture methods.  METHODS: Retinal and visual cortical neurons isolated from new-born rats were firstly cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% new-born calf serum and 10% F12 nutrient mixtures, followed by maintaining culture in Neurobasal medium containing 2% B27 serum-free supplements. Nissls staining was performed for neuron identification. RESULTS AND CONCLUSION: Cultured neurons grew well with plump cell bodies and long processes. Nissls staining showed that the purity of retinal neurons was greater than 90% and the proportion of visual cortical neurons was higher than 50%. The results suggested that there are some differences in culturing methods and growth characteristics of retinal and visual cortical neurons of new-born rats, accordingly, different culture methods are required to obtain high purity neurons. Related Articles | Metrics

Select

Research progress and possible mechanisms of transplantation tolerance induced by mesenchymal stem cells Zhang Ya-yong, Jiang Li-hong, Hou Zong-liu 2010, 14 (14):  2652-2656.  doi: 10.3969/j.issn.1673-8225.2010.14.039 Abstract ( 84 )   PDF (270KB) ( 358 )   Save BACKGROUND: Mesenchymal stem cells are capable of self-renew, a high degree of proliferation, with multi-differentiation potential, low immunogenicity and immunomodulatory properties. Both in vivo and in vitro it is able to regulate allogeneic immune response, to induce immune tolerance. In the solid organ transplantation it is playing an increasingly important role. OBJECTIVE: To summarize the research progress on the immunomodulatory mechanism and application of mesenchymal stem cells in solid organ transplantation. METHODS: An online search of Pubmed was undertaken by using the key words of “Mesenchymal Stem Cells, Mesenchymal Stem Cell Transplantation, Organ Transplantation, Transplantation Immunology, Immunologic Graft Enhancement, Graft vs. Host Disease” in Mesh to identify the relevant articles published in English from January 1994 to October 2009. At the same time, Wanfang database was screened to identify the relevant articles published between January 1994 and October 2009 with the key words of “Mesenchymal Stem Cells, Organ Transplantation, Transplantation Immunology” in Chinese. Inclusive criterion: The articles related to the immunomodulatory properties, transplantation immunology and application of mesenchymal stem cells in the solid organ transplantation were included. Exclusive criterion: The articles with repetitive research or Meta analysis were excluded. RESULTS AND CONCLUSION: Totally 200 relevant articles were selected and 86 of them met the inclusive criterion. Mesenchymal stem cells exhibit low immunogenicity and immunomodulatory properties, have an indispensable advantage about inducing graft tolerance and repairing tissue in solid organ transplantation. The mechanism of inducting immune tolerance may be related to soluble factors, regulatory T cells, tolerant dendritic cells, bone marrow chimerism, anti-inflammatory and tissue repair, dose and time of injecting MSCs. Related Articles | Metrics

Select

Collection of peripheral blood stem cells from two infants of young age and low body mass: Safety and adverse effects Jing Hua, Zhang Jin-Yuan, Lü Xu-jing, Shen Xu-dong 2010, 14 (14):  2657-2660.  doi: 10.3969/j.issn.1673-8225.2010.14.040 Abstract ( 130 )   PDF (251KB) ( 442 )   Save BACKGROUND: The rapidly developed transplantation of peripheral blood stem cells have been successfully used to substitute bone marrow transplantation and become the first choice method for transplantation of hemopoietic stem cells. It is relatively difficult to collect peripheral blood stem cells from young age and low body mass infants. OBJECTIVE: To investigate the safety and adverse effects of peripheral blood stem cell collection from young age and low body mass through the use of blood cell separator. METHODS: Two type 1 diabetes mellitus infants, aged younger than 2 years old and with body mass less than 15 kg, were treated using autologous hemopoietic stem cell transplantation. The two infants were adequately comforted to lesion the fear of collection. At 1 week prior to collection, calcium agent was orally taken to reduce the incidences of low calcium. Within 24 hours prior to collection, oily food was forbidden, and on the day of collection, milk was forbidden, to avoid chylemia, which influences blood collection. Prior to collection, 200 mL 25 Gy γ-ray radiated red blood cells suspension was injected into the tube, which was routinely placed in the subclavian vein, to avoid hypovolaemic syndrome and the effects on hematocrit. Individualized collection parameters were set. During collection, blood circulation volume was 3, 4 times of systemic blood circulation to ensure sufficient total circulation volume. During isolation, the ratio of ACD to whole blood was kept between 1: 11 and 1: 13 to prevent sodium citrate poisoning. RESULTS AND CONCLUSION: Peripheral blood stem cells were successfully collected during first intention in each infant. During collection, stable vital signs but no adverse effects were observed. After collection, mononuclear cells weighted 14.71×108/kg and 18.82×108/kg respectively, and CD34+ cells were about 34.13×106/kg and 32.38×106/kg, respectively in each infant. Therefore, it is feasible to collect peripheral blood stem cells from infants of young age and low body mass under sufficient psychological preparation. Related Articles | Metrics