Comparison of primary vs immortalized murine hepatocytes

Fundamental research and drug development for personalized medicine necessitates cell cultures from defined genetic backgrounds. However, providing sufficient numbers of authentic cells from individuals poses a challenge. Here, we present a new strategy for rapid cell expansion that overcomes current limitations. Using a small gene library, we expanded primary cells from different tissues, donors and species. Cell type specific regimens that allow the reproducible creation of cell lines were identified. In depth characterization of a series of endothelial and hepatocytic cell lines confirmed phenotypic stability and functionality. Applying this technology enables rapid, efficient and reliable production of unlimited numbers of personalized cells. As such, these cell systems support mechanistic studies, epidemiological research and tailored drug development. In these experiments primary murine hepatocytes were compared to immortalized murine hepatocytes with respect to their global gene expression pattern. The global gene expression profile of primary and immortalized hepatocytes (cell line e-mHepA) was analyzed. In addition the murine hepatocyte cell line Hepa1-6 and murine fibroblasts were included as controls (two samples each). The immortalized murine hepatocytes were cultivated on normal tissue cultures plastic (2D samples) and as spheroids (3D samples). The hepatocyte cell line e-mHepA was immortalized by the genes TAg, KLF4, ID3 and Core. Each sample was run in duplicate.