Rat Hepatocyte: Saving Time Increases Viability

Objective: Hepatocyte transplantation is a potential alternative to liver replacement in humans. Several methods for hepatocyte isolation in animal models have been published, many of these require extensive handling and can there-fore compromise the viability and function of the isolated cells. The aim of this study is to isolate excessive amount of rat hepatocytes with high viability in a short time period by modifying the standard isolation method of Seglen. Methods: The hepatocyte isolation was performed using thetwo-step enzymatic method of Seglen in 5 rats. The hepatocyte isolation in the remaining 5 rats was realized using a method modifying the amount of solutions, infusion methods and separa-tion systems shortening process steps. The cells were counted, stained with trypan blue, FDA and PI for viability. Results: We observed an increase in cell count and viability of the hepatocytes in a shorter time period with our modifi ed method compared to the standard two-step enzyme method. We suggest that the increased cell viability is related to the shorter isolation duration. Conclusions: We propose using our modifi ed isolation method in a hepatocyte isolation procedure in which obtaining excess cells with high viability is of critical importance