Hepatocyte selection medium eliminating induced pluripotent stem cells among primary human hepatocytes

摘要

Hepatic insufficiency is a fatal liver disease with a significant decrease in functioning hepatocytes. If hepatocytes could be generated from human induced pluripotent stem(hi PS) cells and transplanted into patients with hepatic insufficiency, the disease may become curable. However, a major limitation to this therapeutic strategy is due to the tumorigenicity of hi PS cells and their ability to form cancer. Current methods for eliminating unwanted hi PS cells use genetic manipulation or reagents that are potentially hazardous for hepatocytes; therefore, revised methods are necessary and anticipated. Glucose and arginine are essential cell culture medium ingredients for the survival of most cells, including hi PS cells. However, hepatocytes can produce its own glucose and arginine through galactokinase and ornithine transcarbamylase, respectively. Therefore, it was hypothesized that unwanted hi PS cells could be eliminated in a medium without glucose and arginine, and supplemented with galactose and ornithine instead. This modified medium has been established as hepatocyte selection medium(HSM). So far, attempts to generate a pure colony of mature hepatocytes from hi PS cells have not been successful. After establishment of co-culture in HSM,primary human hepatocytes survive while hi PS cells die within three days. Our latest results regarding a modification of HSM will be introduced in this manuscript.