用于肝细胞异种移植研究的永生化猪肝细胞的建立与特性分析
Establishment and characterization of immortalized porcine hepatocytes for the study of hepatocyte xenotransplantation.
作者信息
Pan X, Du W, Yu X, Sheng G, Cao H, Yu C, Lv G, Huang H, Chen Y, Li J, Li L J
机构信息
First Affiliated Hospital, College of Medicine, State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Zhejiang University, Zhejiang, China.
出版信息
Transplant Proc. 2010 Jun;42(5):1899-906. doi: 10.1016/j.transproceed.2009.11.043.
BACKGROUND
In light of the critical shortage of donor livers, xenogeneic sources offer the best alternative to human hepatocytes for the treatment of acute liver failure. This study investigated whether a combination of simian virus 40 large T antigen (SV40 LT) and human telomerase catalytic subunit (hTERT) genes could immortalize primary porcine hepatocytes that could reverse acute liver failure (ALF) in rats.
METHODS
We cotransfected SV40 LT and hTERT genes into primary porcine hepatocytes to examine the features of the transfected cell lines. We characterized the potentially therapeutic effect of immortalized porcine hepatocytes in a rat model of ALF induced by 90% hepatectomy.
RESULTS
An immortalized porcine hepatocyte cell line, HepLi, was expanded by >250 passages. HepLi cells maintained the defining characteristics of primary porcine hepatocytes, including porcine albumin secretion, urea production, and diazepam metabolism. Intrasplenic transplantation of HepLi cells significantly improved liver function, and significantly prolonging the survival of rats with ALF.
CONCLUSIONS
Cotransfection of SV40 LT and hTERT immortalized primary porcine hepatocytes without tumorigenicity in vitro. The Immortalized porcine hepatocytes served as a potential cell resource for xenotransplantation.
背景
鉴于供体肝脏严重短缺，异种来源为治疗急性肝衰竭提供了替代人类肝细胞的最佳选择。本研究调查了猿猴病毒40大T抗原（SV40 LT）和人端粒酶催化亚基（hTERT）基因的组合是否能使原代猪肝细胞永生化，从而逆转大鼠急性肝衰竭（ALF）。
方法
我们将SV40 LT和hTERT基因共转染到原代猪肝细胞中，以检测转染细胞系的特性。我们在90%肝切除诱导的大鼠急性肝衰竭模型中，对永生化猪肝细胞的潜在治疗效果进行了表征。
结果
一种永生化猪肝细胞系HepLi传代超过250次。HepLi细胞保持了原代猪肝细胞的特征，包括猪白蛋白分泌、尿素生成和地西泮代谢。脾内移植HepLi细胞显著改善了肝功能，并显著延长了急性肝衰竭大鼠的存活时间。
结论
SV40 LT和hTERT共转染可使原代猪肝细胞在体外永生化且无致瘤性。永生化猪肝细胞可作为异种移植的潜在细胞资源。
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