Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US6458589

Title:Hepatocyte lineage cells derived from pluripotent stem cells
Inventors:Rambhatla, Lakshmi; Redwood City, CA, USA
Carpenter, Melissa K.; Castro Valley, CA, USA
Summary:Described herein is a differentiated cell population which has phenotypic characteristics of liver cells and which is obtained by culturing pluripotent stem cells in the presence of a hepatocyte differentiation agent. Central to the invention is the discovery that when pluripotent stem cells are cultured in the presence of a hepatocyte differentiation agent, a population of cells is derived that has a remarkably high proportion of cells with phenotypic characteristics of liver cells. This invention relates to human embryonic stem cells that are allowed to form embryoid bodies and are then combined with the differentiation agent n-butyrate and optionally supplemented with maturation factors. Further disclosed are methods by which n-butyrate is added to human embryonic stem cells in a feeder-free culture such that a remarkably uniform cell population is obtained which is predominated by cells with morphological features of hepatocytes expressing surface markers characteristic of hepatocytes, and having enzymatic and biosynthetic activity important for liver function. Multiple embodiments of the invention are provided. Therapeutic applications for such an abundant source of cells of the hepatocyte lineage include drug screening and treatments to replenish liver function.
Abstract:It has been discovered that when pluripotent stem cells are cultured in the presence of a hepatocyte differentiation agent, a population of cells is derived that has a remarkably high proportion of cells with phenotypic characteristics of liver cells. In one example, human embryonic stem cells are allowed to form embryoid bodies, and then combined with the differentiation agent n-butyrate, optionally supplemented with maturation factors. In another example, n-butyrate is added to human embryonic stem cells in feeder-free culture. Either way, a remarkably uniform cell population is obtained, which is predominated by cells with morphological features of hepatocytes, expressing surface markers characteristic of hepatocytes, and having enzymatic and biosynthetic activity important for liver function. Since stem cells readily proliferate in culture, this system provides an abundant source of cells of the hepatocyte lineage for a variety of applications, such as drug screening, and replenishing liver function in the context of clinical treatment.
US Patent Website:Click Here for Full Text of Patent
Title Number:US6458589
Application Number:US2000000718308
Date Filed:20/11/2000
Date Published:01/10/2002
Assignee:Geron Corporation, Menlo Park, CA, USA


 
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