Construction and identification of immortalized rat astrocyte cell line expressing enkephalin

【摘要】 <正>Objective:To provide a sound cell source for further ex-vivo gene therapy for chronic pain,we attempt to develop an immortalized rat astrocyte cell line that expresses enkephalin regulated by doxycycline. Methods:Retrovirus infection method was employed to develop an immortalized rat astrocyte cell line that could express enkephalin regulated by doxycycline.The hPPE gene expression level of immoralized astroyte cells(IAC)/ hPPE was detected by RT-PCR,indirect immunofluorescence staining and radioimmunoassay. Results:IAC carrying Tet-on system transfected with preproenkephalin gene could secrete enkephalin that was regulated by doxycycline in a dose-dependent manner and hPPE gene activation could be repeated in on-off-on cycles through administration or removal of doxycycline. Conclusion:An immortalized rat astrocyte cell line that secrete enkephalin under the control of doxycycline is established successfully,which provides a research basis for transgenic cell transplantation for analgesia.更多还原

【Abstract】 Objective:To provide a sound cell source for further ex-vivo gene therapy for chronic pain,we attempt to develop an immortalized rat astrocyte cell line that expresses enkephalin regulated by doxycycline. Methods:Retrovirus infection method was employed to develop an immortalized rat astrocyte cell line that could express enkephalin regulated by doxycycline.The hPPE gene expression level of immoralized astroyte cells(IAC)/ hPPE was detected by RT-PCR,indirect immunofluorescence staining and radioimmunoassay. Results:IAC carrying Tet-on system transfected with preproenkephalin gene could secrete enkephalin that was regulated by doxycycline in a dose-dependent manner and hPPE gene activation could be repeated in on-off-on cycles through administration or removal of doxycycline. Conclusion:An immortalized rat astrocyte cell line that secrete enkephalin under the control of doxycycline is established successfully,which provides a research basis for transgenic cell transplantation for analgesia.更多还原