摘要
目的探讨端粒酶逆转录酶基因转染人胎肝细胞以获得永生化肝细胞的可行性。方法四步灌流法分离人胎肝细胞,携端粒酶逆转录酶基因载体转染人胎肝细胞,光学显微镜观察转染细胞形态变化及增殖能力,免疫组织化学和流式细胞技术检测转染细胞白蛋白表达情况,同时鉴定转染细胞是否具有糖原合成、尿素氮合成等肝细胞功能。结果端粒酶逆转录酶基因转染人胎肝细胞可以重建人胎肝细胞的端粒活性。转染细胞培养5d时呈集落样生长,12d时增殖融合成片状,且具有表达肝脏特异性蛋白、白蛋白和具备糖原合成、尿素氮合成等肝细胞功能。结论采用端粒酶逆转录酶基因转染人胎肝细胞技术有望获得永生化肝细胞,这可能为细胞移植和生物人工肝治疗各种难治性肝病提供新的细胞来源。
Objective To investigate the practicability of accqired persistent hepatocyte on telomerase reverse transcriptase gene transfected human fetal hepatocyte. Methods Human fetal hepatocytes were separated by four-step perfusion method. Telomerase reverse transcriptase gene carrier transfected human fetal hepatocytes were brought. Transfected cell morphology and proliferative capacity were observed by optieal microscope.The albumin expression of transfected cells were detected by immunohistological chemistry and flow cytometry technique. At the same time, transfected cells which weather possesses hepatocyte function in the synthesis of glycogen and urea nitrogen were differentiated. Results Telomerase reverse transcriptase gene transfected human fetal hepatocyte could construct telomere activity of human fetal hepatocyte. Cell culture of transfection presented clustey like growth in 5 days. Transfected cell showed fused patchy growth in 12 days. It possessed expressive liver-specific protein and albumin and had hepatocyte function in the synthesis of glycogen and urea nitrogen. Conclusion Use of telomerase reverse transcriptase gene transfected human fetal hepatocyte technique would help to accqire persistent hepatocyte.It has possibility to use cellular transplantation and bio-artificial liver for the treatment of refractory hepatic diseases and provides a new cellular source.
出处
《岭南现代临床外科》
2008年第1期7-11,共5页
Lingnan Modern Clinics in Surgery
关键词
人端粒酶逆转录酶
转染
人胎肝细胞
生物学特性
Human telomerase reverse transcriptase
Transfection
Human fetal hepatocyte
Biological features