AIM： To explore the effect of ciliary neurotrophic factor （CNTF） on retinal ganglion cell （RGC）-5 induced by hydrogen peroxide （H2O2）. METHODS： After cell adherence, RGC-5 culture medium was changed to contai...AIM： To explore the effect of ciliary neurotrophic factor （CNTF） on retinal ganglion cell （RGC）-5 induced by hydrogen peroxide （H2O2）. METHODS： After cell adherence, RGC-5 culture medium was changed to contain different concentrations of H2O2 from 50 to 150 μmol/L at four time points （0.5, 1, 1.5 and 2h） to select the concentration and time point for H2O2 induced model. Two different ways of interventions for injured RGC-5 cells respectively were CNTF as an addition in the culture medium or recombinant lentiviral plasmid carrying CNTF gene transfecting bone mesenchymal stem cells （BMSCs） for co-culture with RGC-5. RESULTS： Compared to the control group, H2O2 led to RGC-5 death closely associated with concentrations and action time of H2O2 and we chose 125 μmol/L and 2h to establish the H2O2-induced model. While CNTF inhibited the loss of RGC-5 cells obviously with a dose-dependent survival rate. Nevertheless two administration routes had different survival rate yet higher rate in recombinant lentiviral plasmid group but there were no statistically significant differences. CONCLUSION： Both the two administration routes of CNTF have effects on RGC-5 cells induced by H2O2. If their own advantages were combined, there may be a better administration route.展开更多
Hematopoietic stem cells(HSCs)are an attractive target for gene therapy,especially for inherited blood diseases.Moreover,recombinant lentiviral vectors are considered to be prospective in HSCs gene therapy for the hig...Hematopoietic stem cells(HSCs)are an attractive target for gene therapy,especially for inherited blood diseases.Moreover,recombinant lentiviral vectors are considered to be prospective in HSCs gene therapy for the high efficiency of infection.In this study,murine mononuclear cells(MNCs)were isolated from bone marrow and cultured in suspension,and then Lin−CD117+HSCs were isolated by immunomagnetic beads.During culturing,cells and colonies increased in HSCs supplied with cytokines while no change was observed in the control group without cytokines.FUXW recombinant lentiviral vectors were produced by calcium phosphate-mediated transient cotransfection infected MNCs from ICR and C57 mice.The hFIX expressions were 41.7±4.2 ng/mL and 34.5±6.6 ng/mL in supernatant on 7d.The hFIX expressions of HSCs infected by FUXW recombinant lentiviral vectors were 46.6±5.7 ng/mL(with cytokines)and 33.3±4.8 ng/mL(without cytokines)in supernatant on 7d.Results indicate that recombinant lentiviral vectors can infect murine MNCs and Lin−CD117+HSCs efficiently,and expression of the transgene can be improved when supplied with cytokines.展开更多
基金Supported by Ph.D.Programs Foundation of Ministry of Education of China(No.20130141120052)
文摘AIM： To explore the effect of ciliary neurotrophic factor （CNTF） on retinal ganglion cell （RGC）-5 induced by hydrogen peroxide （H2O2）. METHODS： After cell adherence, RGC-5 culture medium was changed to contain different concentrations of H2O2 from 50 to 150 μmol/L at four time points （0.5, 1, 1.5 and 2h） to select the concentration and time point for H2O2 induced model. Two different ways of interventions for injured RGC-5 cells respectively were CNTF as an addition in the culture medium or recombinant lentiviral plasmid carrying CNTF gene transfecting bone mesenchymal stem cells （BMSCs） for co-culture with RGC-5. RESULTS： Compared to the control group, H2O2 led to RGC-5 death closely associated with concentrations and action time of H2O2 and we chose 125 μmol/L and 2h to establish the H2O2-induced model. While CNTF inhibited the loss of RGC-5 cells obviously with a dose-dependent survival rate. Nevertheless two administration routes had different survival rate yet higher rate in recombinant lentiviral plasmid group but there were no statistically significant differences. CONCLUSION： Both the two administration routes of CNTF have effects on RGC-5 cells induced by H2O2. If their own advantages were combined, there may be a better administration route.
基金supported by the National Natural Science Foundation of China (No.30100102).
文摘Hematopoietic stem cells(HSCs)are an attractive target for gene therapy,especially for inherited blood diseases.Moreover,recombinant lentiviral vectors are considered to be prospective in HSCs gene therapy for the high efficiency of infection.In this study,murine mononuclear cells(MNCs)were isolated from bone marrow and cultured in suspension,and then Lin−CD117+HSCs were isolated by immunomagnetic beads.During culturing,cells and colonies increased in HSCs supplied with cytokines while no change was observed in the control group without cytokines.FUXW recombinant lentiviral vectors were produced by calcium phosphate-mediated transient cotransfection infected MNCs from ICR and C57 mice.The hFIX expressions were 41.7±4.2 ng/mL and 34.5±6.6 ng/mL in supernatant on 7d.The hFIX expressions of HSCs infected by FUXW recombinant lentiviral vectors were 46.6±5.7 ng/mL(with cytokines)and 33.3±4.8 ng/mL(without cytokines)in supernatant on 7d.Results indicate that recombinant lentiviral vectors can infect murine MNCs and Lin−CD117+HSCs efficiently,and expression of the transgene can be improved when supplied with cytokines.
