| Construction and identification of immortalized rat astrocyte cell line expressing enkephalin |
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| 作者姓名： | 徐颖 田玉科 田学愎 安珂 杨辉 |
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| 作者单位： | [1]Department of Anesthesiology, Children＇s Hospital ofChongqing University of Medical Sciences, Chongqing 400014,China [2]Department of Anesthesiology, Tongji Hospital of Tong, iiMedical College, Huazhong University of Science andTechnology, Wuhan 430030, China |
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| 基金项目： | This work was supported by the National Natural Science Foundation of China （No. 30170905 ） and Paul Janssen Foundation. |
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| 摘 要： | 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.
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| 关 键 词： | 鉴定方法 细胞 治疗方法 强力霉素 |
| 修稿时间： | 2007-03-09 |
Construction and identification of immortalized rat astrocyte cell line expressing enkephalin |
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| XU Ying,TIAN Yu-ke,TIAN Xue-bi,AN Ke,YANG Hui.Construction and identification of immortalized rat astrocyte cell line expressing enkephalin[J].Chinese Journal of Traumatology(English Edition),2007,10(6):353-356. |
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| Authors: | XU Ying TIAN Yu-ke TIAN Xue-bi AN Ke YANG Hui |
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| Institution: | 1. Department of Anesthesiology, Children's Hospital of Chongqing University of Medical Sciences, Chongqing 400014,China
2. Department of Anesthesiology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China |
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| 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. |
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| Keywords: | Enkephalins Gene therapy Astrocytes Spinal cord injuries |
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