公开(公告)号：US20160046904A1

公开(公告)日：2016-02-18

申请号：US14779017

申请日：2014-04-08

Applicant: NATIONAL INSTITUTES OF BIOMEDICAL INNOVATION, HEALTH AND NUTRITION ,  OSAKA UNIVERSITY

Inventor： Hiroyuki MIZUGUCHI ,  Kenji KAWABATA ,  Kazuo TAKAYAMA ,  Kiyotoshi SEKIGUCHI

IPC: C12N5/071

CPC classification number: C12N5/067 ,  A61L27/3834 ,  C12N2533/52

Abstract: Provided is a method of stably maintaining and culturing “hepatoblast-like cells” generated during a differentiation-inducing process from pluripotent stem cells to hepatocytes. The present invention also provides a culture product obtained by the culture method. The hepatoblast-like cells can be maintained and cultured stably by bringing a laminin into contact with the hepatoblast-like cells. The method of the present invention which uses a laminin makes it possible for the first time to culture, maintain, and proliferate the hepatoblast-like cells. Desired mature cells such as mature hepatocytes and bile duct epithelial cells can be generated in a short time period and can be acquired at a desired timing by maintaining the hepatoblast-like cells. Further, the resulting cultured hepatoblast-like cells were demonstrated to be capable of being induced to differentiate into mature hepatocytes, mature cholangiocytes, bile duct epithelial cells, and the like. Thus, a substantially pure culture product of the present invention can be utilized as a composition for transplantation for use in regeneration of hepatocytes and/or bile duct epithelial cells.

Abstract translation: 提供了一种在分化诱导过程中从多能干细胞向肝细胞稳定维持和培养“成肝细胞”的方法。 本发明还提供通过培养方法获得的培养物。 通过使层粘连蛋白与成肝细胞样细胞接触，可以维持和保持成肝细胞样细胞并稳定培养。 使用层粘连蛋白的本发明的方法使得可以第一次培养，维持和增殖成肝细胞样细胞。 期望的成熟细胞如成熟肝细胞和胆管上皮细胞可以在短时间内产生，并且可以通过维持成肝细胞样细胞在期望的时间获得。 此外，所得培养的成肝细胞样细胞被证明能够诱导分化成成熟肝细胞，成熟胆管细胞，胆管上皮细胞等。 因此，本发明的基本上纯的培养产物可以用作移植用于肝细胞和/或胆管上皮细胞再生的组合物。