Development of a hybrid artificial liver with hepatocytes derived embryonic stem cell for chronic liver failure patients

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We investigated the use of organoid culture technique to mediate hepatocyte differentiation of murine and primate embryonic stem cell. We used our original organoid culture technique, which induce cultured cells to form an organoid in lumen of hollow fibers by centrifugal force.Mouse ES cells formed an organoid inside of hollow fibers. The ES cell proliferated for more than 20 days of culture and high density culture was achieved. The expressions of mRNAs of hepatocyte specific genes were detected at 15 days of culture, and were sustained for more than 30 days of culture. The functions of albumin secretion and ammonia removal were evaluated under hepatocyte differentiation culture condition. The level of these functions of the ES cells per unit volume of cell culture space increased as culture time, and reached the same level of primary mouse hepatocytes.Cynomolgus monkey ES cell was also investigated as a model of primate ES cell. Cynomolgus monkey ES cells formed an organoid and expressed some liver-specific functions such as albumin secretion and ammonia removal. The activity of ammonia removal of ES cells organoid per unit volume of cell culture space was roughly comparable to that of primary hepatocytes and human hepatoblastoma cell lines.In conclusion, our developed novel organoid culture technique directed both mouse and cynomolgus monkey ES cells along a hepatocyte lineage. This culture technique seems to be a promising technique to develop a hybrid artificial liver.