The Establishment Of A Tupaia Hepatocyte Immortalization Method And Characterization Of Tupaia Immortalized Cell Line ITH6.1

Liver diseases are a series of diseases that pose serious health risks to human. And the appropriate animal models are still necessary for liver drug screening and vaccine development. Compared with other commonly used small animal models, such as mice and rats, tree shrew (Tupaia belangeri) is a kind of experimental animal model with potential application value because of its close relationship with human and non-human primate. Due to the similar hepatitis drug targeting gene with human and the susceptibility to some human hepatic pathogens(such as human hepatic virus), tupaia hepatocytes have many similar characteristics with human hepatocytes. However, this experimental model need to be optimized in many aspects. The instability of the experimental results are due to high variability and long experimental period. Meanwhile, the study of primary tupaia hepatocytes(PTH) is restricted often, because the liver is an organ that is at the end of the differentiation, even after a long and laborious operation to get the primary hepatocytes, the cells will die in a few days. It is difficult to carry out the experimental study of gene transfer and gene knock out for a long time with PTH.To our knowledge, there is no tupaia hepatocyte cell line has ever been successfully established. Therefore, the objective of this study is to establish an immortalized tupaia hepatocyte cell line. In this study, an efficient method of immortalizing tupaia hepatocytes is established by transfecting foreign gene into primary tree shrew hepatocytes.Then, the characteristics of immortalized tupaia hepatocytes strain ITH6.1 (immortalized tupaia hepatocyte 6.1) was analyzed.The research showed that ITH6.1 has a strong proliferative capacity and remain relatively stable cell characteristics cultured for over 100 passages. Further analysis on the characteristics of ITH6.1 by gene sequencing, protein expression, biochemical index and so on, the results showed that ITH6.1 has the typical characteristics of tupaia hepatocytes. At the same time, it also suggests that the cell line has good biological safety.The establishment of tupaia immortalized cell lines may reduce the requirement of experimental animal, provide a more stable experimental object.At the same time, it will provide a more convenient and efficient pre-analysis system in vitro for optimizing tree shrew liver disease model.