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1.
AIM: To explore the ex vivo expansion characteristics of the endothelial progenitor cells (EPCs). METHODS: CD34+ cells were selected from umbilical cord blood mononuclear cells (MNC) by MiniMACS system, expanded at the same conditions as that for total MNC, coincubation of CD34+ and CD34- from the same donation for EPCs. In addition, we tested the effect of vessel endothelial growth factor (VEGF) and passage on cell differentiation, expansion kinetics and apoptosis. EPCs were determined and quantified by immunocytochemistry and flow cytometry. RESULTS: Coculture of CD34+ and CD34-,total MNC led to a significant increase in the expansion of CD34+ cells compared with CD34 enrichment (P＜0.05). There was a trend toward decreased apoptosis in cultures when early passage was performed once the linear cord like structures appeared. There was no significant effect on apoptosis between with VEGF and without VEGF group (P＞0.05). These differentiated EPCs were stained positive for CD34+, von Willebrand factor (vWF), KDR, CD31 and incorporate acetylated low-density lipoprotein (LDL). CD34+ and AC133+cells accounted for 68.2%±6.3% (n=6) and 57.2%±9.8% (n=6) of attaching (AT) cells at day 7 of culture, respectively. CONCLUSIONS: Coculture of CD34+ and CD34- or culture of MNC enhances ex vivo expansion of EPCs. Early passage decreases apoptosis rate, VEGF has no significant effect on ex vivo expansion of EPCs.  相似文献   

2.
AN Xiao-hui  CAI Guo-ping 《园艺学报》2003,19(11):1449-1452
AIM: To confirm the existence of the endothelial progenitor cells in human cord blood and to study its differentiation and development process. METHODS: The mononuclear cells in human cord blood were isolated using lymphocyte separation solution. Then the mononuclear cells were cltured in MCDB131 containing 20% fetal bovine serum. The effects of 5 μmol/L dexamethasone, the extract from bovine brain, insulin and hypoxanine on the proliferation and differentiation of the adherent cells were observed. The morphology of the adherent cells were examined twice daily by inverted phase contrast microscope. CD34 and CD14 expression were determined by FACS. Immunohistochemistry was used to confirm the expression of factor Ⅷ. RESULTS: The proliferative endothelial progenitor cells existed within the CD34-adherent mononuclear cells of human cord blood. Dexamethasone and hypoxanine decreased the number of spindle-shaped cells and caudated cells. Bovine brain extract, insulin and FCS enhanced the number of spindle-shaped cells and caudated cells. CONCLUSION: The existence of endothelial progenitor cells within the CD34 - adherent monouclear cells of the human cord blood was observed and these cells were able to differentiate into endothelial-like cells in vitro.  相似文献   

3.
The vascular endothelial progenitor cells are a population of functional endothelial precursors in circulating blood, which are derived from bone marrow or cord blood. CD34+, Flk-1+ and ACl33+ are their molecular markers. In this review, the functional characterization of vascular endothelial progenitor cells is introduced and the relationship between vascular endothelial progenitor cells and angiogenesis in is chemic cardiovascular diseases is discussed. These data may offer a foundation for the development of therapeutic angiogenesis for the prevention and treatment of ischemic cardiovascular diseases by transplantation of vascular endothelial progenitor cells.  相似文献   

4.
AIM: To compare the methods of two currently employed isolation methods for endothelial progenitor cells (EPCs): from total peripheral blood mononuclear cells (PBMCs) and from enriched CD133+ cells, by defining the cell morphology, phenotype, reproductive activities and function in vitro, providing a reference for clinic application. METHODS: PBMCs from the healthy subjects were used for CD133+ sorting or not. The two groups of isolated cells were suspended in complete medium M199 for 7 d to 14 d. EPCs phenotype were characterized by FACS. The proliferation of differentiated EPCs was studied by MTT assay, and VEGF concentration was measured using an ELISA kit. Matrigel experiment and migration assay were imitated vascularization in vivo. RESULTS: PBMCs produced more colony-forming units (CFU) than CD133+ cells from the same volume of blood (P<0.01). From 7 d to 14 d, the two groups show decreased expression of hematopoietic stem cell markers and increased level of endothelial markers, but CD144+ cells in CD133+ group were lower than those in PBMCs groups (P<0.01). Cells in PBMCs group secreted more VEGF than that in CD133+ group on 7 d (P<0.01). Compared to CD133+ group, PBMCs group showed more potential of proliferation and vascularization in vitro. CONCLUSION: CD133+ sorted cells show a lower capacity of differentiation, secretion, proliferation and vascularization in vitro, which is unable to differentiate to mature endothelial cells, indicating that its not a preferential way to obtain EPCs for clinic therapy.  相似文献   

5.
AIM: To direct embryonic stem cells (ESCs) into hematopoietic stem cells (HSCs) in vitro by simulating the hematogenic microenvironment in human early embryonic aorta-gonad-mesonephero (AGM) region.METHODS: Murine E14 embronic stem cell line was used for two-step differentiation.In the first step of primary differentiation,E14 ESCs were seeded into semisolid methylcellulose-based medium containing bone morphogenesis protein 4 (BMP4) and vascular endothelial growth factor (VEGF) for embryoid body (EB) formation.On days 3,6,9,12 and 15,single EB cells were analyzed for Flk-1+ cells amount through flow cytometry.In the second step,single cell from EB containing most Flk-1+ cells was further co-cultured with human AGM stromal cells in non-contact system.On co-culture days of 3,6,9 and 12 days,cells were collected for cell count,flow cytometry for Sca-1+c-kit+ cells analysis,and colony forming cell assay.RESULTS: During the EB formation,BMP4+VEGF promoted Flk-1+ cell genesis on day 9 at peak pencentage value of 27.53%±2.84％,which was statistically higher than that in control group as 8.77±1.10 (P<0.05).Collagenase-disassociated single cell from day 9 EB was co-cultured with human AGM stromal cells of hAGMS3 or hAGMS4 for further hematopoietic differentiation.On day 6 Sca-1+c-kit+ cells got to peak value as 7.31%±1.21％ ［(2.57±0.48) folds］ and 7.62%±1.52％ ［(2.35±0.36) folds］ in hAGMS3 and hAGMS4 feeder systems,respectively,both of which were greater than those values of no-stroma groups at the same culture duration (P<0.05).Colonogenic cell assay showed that these Sca-1+c-kit+ cells had ability of forming multiple lineage hematopoietic colonies.CONCLUSION: BMP4 in combination with VEGF promotes Flk-1+ cell genesis during EB formation in vitro.Stromal cells from early human embryonic AGM region further enhance the directed differentiation of these primitive cells into HSCs.This two-step induction differentiation model can be used for molecular mechanism study of ESCs hematopoietic differentiation.  相似文献   

6.
AIM:To investigate the expression and function of homing related molecules and transmigration ability of human cord blood CD34+ hematopoietic stem/progenitor cells after short time stimulation with cytokine SCF and IL-6.METHODS:CD34+ cells were separated by Ficoll density gradient centrifugation and stimulated by SCF and IL-6 cytokines for 48 h. The changes of CD49d (VLA-4), CD11a (LFA-1), CD62L (L-selectin) and CD184 (CXCR4) were analyzed by flow cytometry. The adherent and migration activities of CD34+ cells were evaluated in human fibronectin (FN) coated microplates (96 wells) and transwell system.RESULTS:The numbers of CD34+ cell expanded to 3 folds and the percentages of CD34+ cells that were positive expressions for CD49d, CD11a, CD62L or CD184 increased 1 to 2 folds after the cytokine stimulation. The spontaneous adhesion between CD34+, FN and SDF-1 induced migration increased after SCF+IL-6 stimulated.CONCLUSION:SCF+IL-6 can improve the most of the homing related characteristics and activities in the short time expansion of CD34+ hematopoietic stem/progenitor cells, which may be partly related to the increased intrinsic homing potential.  相似文献   

7.
AIM:To study the effect of proliferating cell nucler antigen antisense oligonucleotide on ex vivo expansion of cord blood CD34+ hematopoietic stem/progenitor cells. METHODS:CD34+ cells were purified from fresh cord blood by immunomagnetic beads. CD34+ cells were incubated in liquid culture system with different concentrations of PCNA-ASODN. Using flow cytometry, the number of different kinds of stem/progenitor cells and PCNA expression were measured after CD34+ cell incubation. RESULTS:PCNA was lowly expressed in low experiential group, with a positive rate of (27.2±3.6)% and (19.0±1.5)%, the positive rate of control group was (53.8±8.3)% (P<0.01), a high significant difference was observed. Just in low concentration of PCNA-ASODN, the percentage of CD34+cells were increased to (33.4±3.2)%, CD34+CD38-cells expanded (57.8±9.9) folds, and the percentage of CD34+cells were (25.2±2.6)% (P<0.01), the CD34+CD38-cells expanded (43.5±7.4) folds (P<0.05), it has significant difference compared with the control group. CONCLUSION:Low concentration of PCNA-ASODN decreases PCNA expression effectively, slows down differentiation of CD34+cells during ex vivo expansion procedure, and improves the expansion efficiency.  相似文献   

8.
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9.
AIM:To purify murine yolk sac endothelial cells (mYS-EC) and investigate the cytokines mRNA expression in mYS-EC. METHODS:The murine yolk sacs were digested with 0.1% collagenase, resuspended in DMEM and counted after digestion and centrifugation. The yolk sac adherent cells were cultured in DMEM containing 15% FBS with 10% mBMEC-CM or 5μg/L VEGF, ECGF and bFGF. The phagocytose function and expression of vWF were evaluated via particle phagocytosis and immunohistochemistry method. Atlas cDNA expression array was used for analysis of cytokine expression in mYS-EC. RESULTS:Colonies consisting of pure yolk sac endothelial cells were obtained in liquid culture system containing 15% FBS and 10% mBMEC-CM or 5μg/L VEGF, ECGF and bFGF. For complete purification of the endothelial cells, subsequent passage was also necessary. Cellular cord formed during passage culture. The endothelial cells were round or oval sharp in morphology, positive in phagocytosis and factor VIII related antigen (von Willebrand's Factor, vWF). The mRNA expressions of cytokines, such as TGF-β2, TNF-α, IFN-γ, FL, BMP-4, MIP-1β, BMP-2A, FLT2, endothelin 2, thymosin β10, IL-6, IL-13, IL-9, SCYA5 and ACBP were detected in mYS-ECs. CONCLUSION:mYS-EC was purified and expanded in vitro. The mRNA expression of 15 kinds of cytokines was detected in mYS-ECs by Atlas arrays.  相似文献   

10.
AIM: To determine the combined effect of transmuscle laser revascularization (TMR) and endothelial progenitor cells(EPCs) treatment on ischemic hindlimb of nude rats.METHODS: Mononuclear cells (MNCs) isolated from human umbilical cord-blood (HUCB) by density gradient centrifugation were expanded in vitro. Immunocytochemistry and flow cytometry studies were performed. EPCs were labeled with 1, 1- dioctadecyl-1 to 3, 3, 3, 3- tetramethyl-indocarbocyanine perchlorate (DiI) before injected into the laser induced channels or ischemic region. Acute ischemic limb was created in 4 groups of nude rats by ligating right external iliac artery. All animals were divided randomly into the following four groups: TMR+EPCs group: local transplantation of EPCs into laser channels; TMR group: transmuscular channels were created without EPCs; EPCs group: EPCs were injected into ischemic hindlimb; control group: ischemic model without TMR or EPCs. All rats underwent femoral artery ultrasonic blood flow measurements of the ischemic and nonischemic limbs to obtained a flow ratio ［femoral artery flow index (FAFI): right femoral artery flow /left femoral artery flow］ at baseline (after ligating artery immediately) and 28 days postoperation, and then the samples of ischemic limb muscle underwent histochemical and immunohistologic analysis. RESULTS: The attached cells expressed endothelial cell (ECs) markers (KDR, CD34, CD31, AC133 and von Willebrand factor) and exhibited function similar to that of ECs judged by Ac-LDL incorporation. Flow cytometric analysis disclosed that AT cells were positive for CD34 (62%±7%) and AC133 (57.2%±9.8%) at day 7 of culture. 28 days after therapy, FAFI was significantly higher in the TMR +EPCs (0.66±0.09, P<0.01) and EPCs group (0.59±0.09, P<0.05) compared to control group (0.47±0.05). It was significantly higher in TMR +EPCs-，EPCs- and TMR group compared to baseline (TMR+EPCs group: 0.66±0.09 vs 0.39±0.07, P<0.01; TMR group: 0.54±0.12 vs 0.40±0.09, P<0.05; EPCs group: 0.59±0.09 vs 0.38±0.08, P<0.01; control group: 0.47 ±0.05 vs 0.39±0.08, P>0.05). FAFI in the control group was unchanged and no difference was found between TMR group and control group. TMR+EPCs (5.66±0.77), TMR (4.96±0.31) as well as EPCs (4.68±0.44) treatment resulted in an increased number of capillaries in the treated regional area compared with control group (2.60±0.31, P<0.01).CONCLUSION: Nd: YAG-laser revascularization combined with the application of EPCs transplantation significantly ameliorates perfusion and augments neovascularization in this ischemic hindlimb model.  相似文献   

11.
AIM: To investigate the differentiation potential of human placenta derived mesenchymal stem cells (PDMSCs) into endothelial cells (ECs) in vitro. METHODS: PDMSCs were isolated from human placenta tissues, characterized by flow cytometry and induced to differentiate into endothelial cells with 50 μg/L VEGF and 10 μg/L bFGF. To detect the specific markers of ECs during the process of differentiation, the method of immunocytochemistry was performed. The specific structure and function of endothelial cells were observed by transmission electron microscopy and in vitro angiogenesis assay kit, respectively. RESULTS: CD105 and CD106 were positive in PDMSCs, while CD34,CD45 and CD31 were negative.The ECs differentiated from PDMSCs showed cobblestone-like morphology, and expressed early endothelial marker of Flk-1/KDR and mature endothelial markers of CD31, vWF and CD144/VE-cadherin in a time-dependent manner during the endothelial cell differentiation (0 day, 4 days, 8 days and 12 days). The endothelial specific structure, Weibel-palade body, was observed under transmission electron microscope. The inoculation of ECs on the extra cellular matrix gel formed capillary-like structures. CONCLUSION: Plentiful PDMSCs can be isolated from placenta, and differentiate into the cells with functional characteristics of ECs in vitro, indicating that the placenta tissues will become optimal source of seed cells for vascular engineering and regenerative medicine.  相似文献   

12.
AIM: Endothelial progenitor cells (EPCs) are a group of stem cells/progenitor cells, which exist in postnatal body and can be of specially homing to the foci of angiogenesis and then differentiate into endothelial cells.This investigation was to study the method for culturing endothelia progenitor cells (EPCs) in vitro, and to observe its feasibility and condition formed vessel-like structure.METHODS: The cells were isolated from born marrow, peripheral blood, umbilical cord blood or spleen in different laboratories.The EPCs derived from human umbilical cord blood and rabbit peripheral blood were cultured in vitro through adhesion selection and were differentiated into endothelial cells under the induction of special cytokines.The expression of CD34, VEGFR-2, AC133 and VE-cadherin were detected by fluorescence-activated cell sorting.The endothelial cell lineage was confirmed by DiI-ac-LDL up-taking and Ⅷ factor immunocy tochemistry.RESULTS: The EPCs derived from human umbilical cord blood and rabbit peripheral blood were cultured in vitro successfully, forming cord-like and tube-like structure.The EPCs derived from rabbit peripheral blood differentiated more mature and formed vessel-like structure.CONCLUSION: The EPCs derived from human umbilical cord blood and rabbit peripheral blood formed vessel-like structure in vitro.EPCs may be a potential resource of vessel tissue engineering.  相似文献   

13.
AIM: To investigate the effect of vascular endothelial growth factor(VEGF) on the biological function of peripheral endothelial progenitor cells (EPCs) and to find the suitable concentration to promote the growth of EPCs.METHODS: Total mononuclear cells (MNCs) were isolated from peripheral blood by density gradient centrifugation,and then the cells were plated on fibronectin-coated culture dishes.After culture for 4 days,attached cells were incubated with VEGF in a series of concentrations (0,10,20 and 50 μg/L) for 72 h,then attached cells were characterized with immunohistochemistry.EPC proliferation and migration activity were assayed with MTT assay and modified Boyden chamber assay,respectively.EPC adhesion assay was performed by replating MNCs on fibronectin-coated dishes,and then the adherent cells were counted.RESULTS: The EPCs from MNCs were successfully isolated and were differentiated to endothelial cells (ECs).Incubation of isolated human MNCs with VEGF increased the proliferative,migratory and adhesive capacities of EPCs,and this effect was most prominent when the concentration of VEGF was 20 μg/L after 72 hours.At the same concentration of VEGF,the functions of EPCs from patients with masculine coronary arteriography were lower than those of EPCs from patients with negative coronary arteriography.CONCLUSION: Functional activities of EPCs are decreased in patients with masculine coronary arteriography.The results suggest that the low concentration of VEGF may improve functional activities of EPCs.  相似文献   

14.
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15.
AIM: To probe the effect of different panel reactive antibody(PRA) serum levels from patients with β-thalassemia on the proliferation and differentiation potential of the hematopoietic stem /progenitor cell of cord blood.METHODS: 1×105 mononuclear cells (MNCs) isolated from umbilical cord blood were incubated with different PRA serum levels (0 μL,50 μL,100 μL) respectively and complement,inoculated into the methylcellulose cultural system.The proliferation and differentiation potential of the hematopoietic stem /progenitor cell of cord blood by the colony formation assay were detected on day 7 and day 14,respectively.RESULTS: After culture of 7 days,the total colonies and CFU-GM were 88.20±9.41,79.00±11.39 in group A and 88.60±9.12,79.20±10.44 in group B,which were significantly higher than those of 20.60±7.39,15.20±4.66 in group C and those of 4.00±2.05,1.40±0.51 in group D (P<0.01).Meanwhile compared with group A and group E,no significant difference was found (P>0.05).After culture for 14 days,the total colonies and CFU-GM were 216.00±31.10,117.40±24.80 in group A and 213.20±31.06,116.00±19.75 in group B,which were significantly higher than those of 97.80±14.43,32.80±8.10 in group C and those of 31.40±13.41,8.40±4.30 in group D (P<0.01).The CFU-GEMMs were 45.60±8.51 in group A and 42.60±7.03 in group B,which were significantly higher than those of 20.80±6.96 in group C and those of 7.80±6.06 in group D (P<0.05).The BFU-MK was 12.80±4.42 in group A and 11.00±2.74 in group B respectively,which were significantly higher than that of 1.00±0.55 in group D (P<0.05).The CFU-E in group B 17.20±4.03 was significantly higher than that of 5.60±2.87 in group D (P<0.05).Meanwhile compared with group A and group E,no significant difference was found (P>0.05).By the Kendall test,there were negative correlations between the level of PRA serum and the total colonies,CFU-GM on day 7,the total colonies,CFU-GM,CFU-GEMM,BFU-E,BFU-MK on day 14 (tau-b=-0.793,-0.849,-0.808,-0.804,-0.645,-0.674,-0.624,P<0.01).There was a negative correlation between the level of PRA serum and CFU-MK on day 14 (tau-b=-0.466，P<0.05).CONCLUSION: PRA sera inhibit the colony in the colony cultures of the hematopoietic stem cells/progenitor cells in cord blood.The inhibition depends on the level of PRA sera.The higher the level of PRA sera,the stronger the inhibition is observed in our study.  相似文献   

16.
AIM: To explore the feasibility and biological characterization of long-term regulated expansion of JAK2 transduced human CD34+ cord blood cells in vitro.METHODS: A retrovirus (RV) vector which contains JAK2 catalytic domain and two binding sites for a chemical inducer,dimerization (AP20187),was cloned (designated MGI-F2JAK2).CD34+cells were enriched from cord blood with a MiniMACS system.The purified CD34+ cells were transfected with supernatant from the retrovirus packaging cell line that expressed JAK2.Following transduction,cells were expanded into four groups: AP20187 alone,FL alone,TPO,alone,AP20187+FL+TPO,respectively.The expanded cells were monitored by GFP expression,immunophenotyping,progenitor colony assay,karyotype analysis as well as tumorigenesis in nude mice.RESULTS: The purity of selected CD34+ cells was over 91% and gene transfer rate was 49.32%±6.21%.Only the group of AP20187 +FL+ TPO was obtained a significant sustained outgrowth of the transduced CD34+ cord blood cells.The percentage of GFP+ cells consistently produced a rise to the 90% peak level by the end of 8th week of culture.Flow cytometry analysis showed that the phenotype of the expanded cells was CD33+,CD61+ and Gly-A+ partial positive;CD38+ and HLA-DR+ strong positive,while CD2,CD7 and CD19 were almost negative.Colony assays performed in methycelluos,which can give rise to BFU-E,CFU-GM and CFU-Mix,the CFU-GM was predominantly in all colonies.The tumor was not observed in nude mice and the karyotype analysis was normal from expanded cells.CONCLUSION: The results demonstrate that AP20187-mediated activation of JAK2 signaling is capable of stimulating expansion JAK2 transduced CB CD34+ cells in combination with FL and TPO.This system may have applications for studies in signaling transduction,hematopoiesis,and for gene and cell therapy.  相似文献   

17.
AIM: To investigate the effects of interleukin-6 (IL-6) and interleukin-11 (IL-11) on differentiation of cord blood CD34+ cells towards megakaryocytes and platelet production in vitro.METHODS: The CD34+ cells from fresh umbilical cord blood samples were cultured in serum-free culture medium with thrombopoietin (TPO) 50 μg/L,IL-3 10 μg/L,stem cell factor (SCF) 50 μg/L as control groups,then 10 μg/L IL-6 or IL-11 or IL-6＋IL-11 respectively was added as treatment groups.Mononuclear cells (MNCs) in cultured cells were detected by cell counter,megakaryocytes (CD41+ cells) and platelets were measured by flow cytometry,respectively.Platelet agglutination after thrombin induced was observed by microscopy and flow cytometry.RESULTS: Compared with the control group,the number of MNCs was not significantly different（P>0.05）,but the numbers of CD41+ cells and platelets were increased significantly (P<0.05) in treatment groups.There were more platelet particles in treatment groups than those in control group by microscopy and the results also showed that the cytoplasmic fragments from the cultures responded to thrombin induction.CONCLUSION: It is concluded that both IL-6 and IL-11 induce the cord blood CD34+ cells to differentiate towards megakaryocytes and produce platelets.  相似文献   

18.
AIM: Humanized-NOD/SCID(hu-NOD/SCID) mouse model was established and the level of immune reconstitution was assessed in this model. METHODS: Mononuclear cells (MNC) and CD34+ cells were isolated or sorted from cord blood(CB). Human CD45, CD19, CD3 markers on cells from NOD/SCID murine peripheral blood(PB), bone marrow(BM), thymus were detected by FCM from 4 to 10 weeks after hematopoietic stem cell transplantation. After 10 weeks, the gene expressions of the human β2M and RAG2 were detected by RT-PCR in PB or bone marrow of mice model. RESULTS: Human CD45, CD19, CD3 cells populations in PB and BM were found by flow cytometry in mice model transplanted with CD34+ cells or CB MNC from 4 to 10 weeks. The highest positivity of human lymphocytes was at 8 week after transplantation. The levels of human cell engraftment in mice transplanted with CD34+ cells were higher than those in mice transplanted with CB MNC. The mRNA of human β2M and RAG2 were found by RT-PCR in BM.CONCLUSION: The higher level of human lymphocyte engraftment is established in NOD/SCID mouse model transplanted with CD34+ compared with CB MNC. The maturation of T lymphocytes could be happened in bone marrow of mice model.  相似文献   

19.
AIM: To study the effects of VEGF over-expressed C6 glioma cells on the expression of Flk-1and Flt-1in cocultured microvascular endothelial cells using the rat hepatic cells BRL 3A as control. METHODS: Cocultured systems of rat pulmonary microvascular endothelial cells with C6 and endothelial cells with BRL 3A were established. Immunocytochemical method was used to investigate the expression change of Flk-1and Flt-1protein in cocultured microvascular endothelial cells. The expression of Flt-1and Flk-1mRNA was analyzed by RT-PCR and Northern blot. RESULTS: The microvascular endothelial cells cocultured with C6 showed increased expression of Flk-1and Flt-1protein (P <0.05), while that cocultured with BRL 3A, showed decreased expression of Flt-1and Flk-1protein(P <0.01). The results of RT-PCR and Northern blot showed that the Flk-1, Flt-1mRNA of microvascular endothelial cells were markedly up-regulated after coculture with C6 glioma cells (P <0.01), while the endothelial cells cocultured with BRL 3A had down-regulated expression of Flk-1, Flt-1mRNA (P <0.01). CONCLUSION: VEGF over-expressed C6 glioma cells may markedly up-regulate the expression of Flt-1and Flk-1in cocultured microvascular endothelial cells. These results suggest that this effect could be one of the important mechanisms in glioma angiogenesis in vivo.  相似文献   

20.
AIM：To observe the growth pattern and surface markers of human umbilical cord mesenchymal stem cells(hUCMSCs) and to explore the influence of basic fibroblast growth factor (bFGF) on the proliferation and collagen production in hUCMSCs cultured in vitro. METHODS：hUCMSCs were isolated by enzyme digestion method and adherent culture. The surface markers CD45, CD34, CD105, CD29 and HLA-DR of the cells were analyzed by flow cytometry. The osteogenic ability and adipogenic differentiation were confirmed with oil red O and alizarin red staining. The optimal concentration of bFGF to promote the proliferation of hUCMSCs was 20 μg/L. The cells in control group were cultured in the growth medium consisting of DMEM/F12 and 10% volume fraction of fetal bovine serum. The cells in experiment group were cultured under the same condition of control group but plus 20 μg/L bFGF. The proliferation of hUCMSCs was analyzed by MTT assay. The expression of type I and III collagens at mRNA and protein levels was determined by RT-PCR and Western blotting. RESULTS：The growth curves indicated that bFGF promoted the proliferation of hUCMSCs. The hUCMSCs expressed CD29, but did not express CD34, CD45 or HLA-DR in the presence or absence of bFGF unanimously. The cells were alizarin red staining-positive and oil red O staining-positive. Compared with control group, the expression of type I and III collagens significantly decreased at mRNA and protein levels in experiment group. CONCLUSION：bFGF promotes the proliferation of hUCMSCs and does not change the expression of the surface markers. bFGF inhibits the expression of type I and III collagens at mRNA and protein levels, indicating that bFGF enhances the healing of wound without inducing scar hyperplasia.  相似文献   

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