Construction and identification of immortalized rat astrocyte cell line expressing enkephalin.
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.


CONCLUSIONS
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.
Department of Anesthesiology, Children's Hospital of Chongqing, University of Medical Sciences, Chongqing 400014, China.
Ying
Y
Xu
Yu-ke
YK
Tian
Xue-bi
XB
Tian
Ke
K
An
Hui
H
Yang
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001253:Astrocytes; D002461:Cell Line, Transformed; D002908:Chronic Disease; D004318:Doxycycline / Q000494:pharmacology; D004745:Enkephalins / Q000235:genetics* / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D018377:Neurotransmitter Agents / Q000235:genetics* / Q000378:metabolism; D059408:Pain Management; D011498:Protein Precursors / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D012190:Retroviridae / Q000235:genetics
D000900:Anti-Bacterial Agents; D004745:Enkephalins; D018377:Neurotransmitter Agents; D011498:Protein Precursors; C033660:preproenkephalin; D004318:Doxycycline