Transplantation of hepatocyte-like cells differentiated from ES cells

1)胚胎干细胞诱导肝细胞的研究我们已经成功地用胚胎干细胞的EB诱导出了产白蛋白细胞。这些细胞有生产尿素的能力。综上所述，这些细胞被认为是肝细胞。将该肝细胞样细胞预先移植到肝损伤小鼠C57BL/6体内。移植细胞在受体肝脏中生长并产生白蛋白。但20-30%小鼠出现畸胎瘤形成。选择细胞是为了抑制畸胎瘤。首先，对EB进行单层细胞培养，并进行分离。用ercoll法比重力法筛选细胞。该细胞组于10月3 - 4日呈阴性。当将该细胞移植到上述肝损伤模型小鼠时，未发现畸胎瘤形成。如上所述，从胚胎干细胞中获得了不产生畸胎瘤的细胞群的恢复。此外，通过使用FAGS和MACS去除该细胞组的其他血细胞成分，可以获得许多白蛋白阳性细胞。2)食蟹猴胚胎干细胞针对胚胎干细胞的临床应用，尝试用食蟹猴胚胎干细胞诱导肝细胞。虽然需要采用不同的小鼠胚胎干细胞诱导方法来诱导食蟹猴干细胞，但我们的方法能够诱导产生白蛋白的细胞。3)脐带血诱导肝细胞我们成功地用脐带血诱导肝细胞。识别出两种细胞类型，一种是表达白蛋白和CK-18，另一种是表达白蛋白和CK-19。目前，我们正在研究脐带血诱导胆管上皮细胞。

1)A study of induction of hepatocytefrom ES cellWe have succeeded in inducing albumin-producing cell through EB from ES cell. These cell had a ability of producing urea. Taken these together, these cells are considered to be hepatocyte. This hepatocyte-like cell was transplanted into C57BL/6 mouse which had liver injury in advance. The transplanted cell grew up in the recipient liver and produced albumin. But, teratoma formation was seen in 20-30% mouse. The cell was selected in order to suppress teratoma. First, the single layer cell culture from EB was performed, and isolated. The cell was selected by specific gravity by the ercoll method. This cell group was negative in oct 3/4. When this cell was transplanted to an above-mentioned liver injury model mouse, teratoma formation was not recognized. As mentioned above, recovery of the cell group which does not generate teratoma from an embryonic stem cell was attained. Furthermore, it became possible by removing other blood cell ingredients for this cell group using FAGS and MACS to obtain many albumin positive cells.2) ES cell of cynomolgus monkeyAiming at clinical application of embryonic stem cell, induction of a liver cell was tried from cynomolgus monkey embryonic stem cell. Although a different method of induction from a mouse embryonic stem cell was required for induction of cynomolgus monkey stem cell, albumin-producing cell was able to be induced by our method.3) Induction of hepatocyte from umbilical cord bloodWe succeeded in liver cell induction from umbilical cord blood. Two cell types were recognized, one is expressing albumin and CK-18 and the other is expressing albumin and CK-19. Now, we are studying bile duct epithelium cell induction from umbilical cord blood.