Jiang L, Li J G, Lan L, Wang Y M, Mao Q, You J P
Institution of Infectious Diseases, Southwest Hospital, Third Military Medical University of the Chinese People’s Liberation Army, Chongqing, China.
Transplant Proc. 2010 Nov;42(9):3773-8. doi: 10.1016/j.transproceed.2010.08.064.
Chimeric mice with repopulated human hepatocytes are widely used for drug development research. HepaRG cell line is a naturally immortalized human liver cell line with progenitor properties and inducible bipotent differentiation capability. It would be useful if HepaRG cells could repopulate damaged livers severe, combined immunodeficient × beige (SCID/bg) mice and exhibit special human hepatic features. After inducing mouse hepatocyte apoptosis with an antimouse agonistic Fas monoclonal antibody (Jo2 mAb), HepaRG cells with bipotent differentiation capability were repopulated in SCID/bg mouse livers. HepaRG cells were transplanted intrasplenically into SCID/bg mice treated with 0.2 mg/kg Jo2 mAb once a week for 10 weeks. Human hepatocyte repopulation was characterized by immunohistochemistry for human serum albumin (HSA), Hep Par1, and CK18 and by immunofluorescence staining for HSA. Human albumin mRNA was detected by reverse-transcription polymerase chain reactions. HSA levels were quantified by Western blotting and enzyme-linked immunosorbent assays. Our results showed that HepaRG cell engraftment protected mice from the effects of Jo2 mAb-mediated liver hemorrhage and hepatocyte apoptosis. At 2 weeks after transplantation, increase concentrations of HSA were detected in recipient blood and liver. At 12 weeks after transplantation, approximately 15%-20% of mice showed livers repopulated with human hepatocytes. In conclusion, normal SCID/bg mice were suitable recipients for HepaRG cell transplantation when induced with Jo2 mAb. This chimeric mouse model with HepaRG cells could serve as a useful model for in vivo studies of drug metabolism and hepatitis virus infections.
具有重新填充人肝细胞的嵌合小鼠被广泛用于药物开发研究。HepaRG细胞系是一种具有祖细胞特性和可诱导的双能分化能力的天然永生化人肝细胞系。如果HepaRG细胞能够在严重联合免疫缺陷×米色（SCID/bg）小鼠的受损肝脏中重新填充并表现出特殊的人肝特征，那将是很有用的。在用抗小鼠激动型Fas单克隆抗体（Jo2 mAb）诱导小鼠肝细胞凋亡后，具有双能分化能力的HepaRG细胞在SCID/bg小鼠肝脏中重新填充。将HepaRG细胞经脾内移植到每周用0.2 mg/kg Jo2 mAb处理10周的SCID/bg小鼠体内。通过人血清白蛋白（HSA）、Hep Par1和CK18的免疫组织化学以及HSA的免疫荧光染色来表征人肝细胞的重新填充情况。通过逆转录聚合酶链反应检测人白蛋白mRNA。通过蛋白质免疫印迹法和酶联免疫吸附测定法定量HSA水平。我们的结果表明，HepaRG细胞植入可保护小鼠免受Jo2 mAb介导的肝出血和肝细胞凋亡的影响。移植后2周，在受体血液和肝脏中检测到HSA浓度升高。移植后12周，约15%-20%的小鼠肝脏显示有人肝细胞重新填充。总之，当用Jo2 mAb诱导时，正常的SCID/bg小鼠是HepaRG细胞移植的合适受体。这种具有HepaRG细胞的嵌合小鼠模型可作为药物代谢和肝炎病毒感染体内研究的有用模型。
