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| 携带SV40T/HSV-tk基因永生化正常成人肝细胞的构建 | |
| 引用本文： | 姚文芳,崔忻,田磊,高宇红,洪权,薛毅珑. 携带SV40T/HSV-tk基因永生化正常成人肝细胞的构建[J]. 军医进修学院学报, 2011, 32(11): 1139-1141. DOI: 11-3275/R.20110627.0929.002 |
| 作者姓名： | 姚文芳 崔忻 田磊 高宇红 洪权 薛毅珑 |
| 作者单位： | 1. 解放军总医院老年医学研究所细胞生物学研究室,北京100853;首都医科大学附属北京天坛医院超声科,北京100050 2. 解放军总医院老年医学研究所细胞生物学研究室,北京,100853 3. 解放军总医院全军肾病中心暨重点实验室,北京,100853 |
| 基金项目： | 国家“863”计划项目 |
| 摘 要： | 目的构建永生化正常成人肝细胞为生物型人工肝提供安全有效的人肝细胞源。方法转染包装细胞,获得逆转录病毒;将该病毒感染正常成人肝细胞HL-7702;更昔洛韦毒性实验检测转染后肝细胞内HSV-tk基因;RT-PCR法检测转染后细胞内SV40T的mRNA;Western Blot法和免疫荧光染色检测转染后细胞内SV40T基因的蛋白表达。结果更昔洛韦检测转染后的肝细胞存活良好,而转染前的肝细胞死亡。RT-PCR法显示转染后的肝细胞内存在SV40T mRNA;Western Blot法和免疫荧光染色法显示转染后肝细胞内存在SV40T蛋白。结论转染后的正常成人肝细胞中含有SV40T和HSV-tk基因,并表达相关蛋白。 |
| 关 键 词： | 永生化 正常成人肝细胞 逆转录病毒 自杀基因 |
Construction of immortalized human liver cells with SV40T/HSV-tk gene |
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| YAO Wen-fang,CUI Xin,TIAN Lei,GAO Yu-hong,HONG Quan,XUE Yi-long. Construction of immortalized human liver cells with SV40T/HSV-tk gene[J]. Academic Journal of Pla Postgraduate Medical School, 2011, 32(11): 1139-1141. DOI: 11-3275/R.20110627.0929.002 | |
| Authors: | YAO Wen-fang CUI Xin TIAN Lei GAO Yu-hong HONG Quan XUE Yi-long |
| Affiliation: | 1Cell Biology Laboratory,Institute of Gerontology,Chinese PLA General Hospital,Beijing 100853,China;2Chinese PLA Center of Nephrology and Key Laboratory,Chinese PLA General Hospital,Beijing 100853,China;3Department of Ultrasound,Beijing Tiantan Hospital,Beijing Medical University,Beijing 100050,China |
| Abstract: | Objective To provide safe and effective artificial liver human liver cells by constructing immortalized normal adult liver cells.Methods Artificial liver human liver cells were transfected and packaged to obtain retrovirus which was used to infect the normal adult liver cells HL-7702.Liver cells with the HSV-tk gene were detected by ganciclovir toxicity test.SV40T mRNA-transfected liver cells were detected by RT-PCR.SV40T gene expression in transfected liver cells was detected by Western blot with immunofluorescence staining.Results Ganciclovir toxicity test showed that the transfected liver cells survived well while the untransfected liver cells died.Immortalized SV40T mRNA was detected by RT-PCR in the transfected liver cells,while SV40T protein was detected by Western blot in the transfected liver cells with immunofluorescence staining.Conclusion SV40T and HSV-tk genes can be detected in transfected normal adult liver cells and express their associated proteins. |
| Keywords: | immortalized normal adult liver cells retroviral suicide gene |
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