Efficient differentiation of functional hepatocytes from human embryonic stem cells.

نویسندگان

  • Sadhana Agarwal
  • Katherine L Holton
  • Robert Lanza
چکیده

Differentiation of human embryonic stem cells (hESCs) to specific functional cell types can be achieved using methods that mimic in vivo embryonic developmental programs. Current protocols for generating hepatocytes from hESCs are hampered by inefficient differentiation procedures that lead to low yields and large cellular heterogeneity. We report here a robust and highly efficient process for the generation of high-purity (70%) hepatocyte cultures from hESCs that parallels sequential hepatic development in vivo. Highly enriched populations of definitive endoderm were generated from hESCs and then induced to differentiate along the hepatic lineage by the sequential addition of inducing factors implicated in physiological hepatogenesis. The differentiation process was largely uniform, with cell cultures progressively expressing increasing numbers of hepatic lineage markers, including GATA4, HNF4alpha, alpha-fetoprotein, CD26, albumin, alpha-1-antitrypsin, Cyp7A1, and Cyp3A4. The hepatocytes exhibited functional hepatic characteristics, such as glycogen storage, indocyanine green uptake and release, and albumin secretion. In a mouse model of acute liver injury, the hESC-derived definitive endoderm differentiated into hepatocytes and repopulated the damaged liver. The methodology described here represents a significant step toward the efficient generation of hepatocytes for use in regenerative medicine and drug discovery.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Expression of Endoderm and Hepatic Specific Genes after in vitro Differentiation of Human Embryonic Stem Cells

Background: Human embryonic stem cells (hESC), which are derived from the inner cell mass of the blastocysts, have been considered to be pluripotent cells. In this study we examine the differentiating potential of hESC into hepatocytes by characterization of the expression of endoderm and liver-specific genes. Methods: hESC were cultivated in suspension to form aggregates, the embryoid bodies. ...

متن کامل

Polarisation and functional characterisation of hepatocytes derived from human embryonic and mesenchymal stem cells.

Adult hepatocytes are polarised with their apical and basolateral membranes separated from neighbouring cells by tight junction proteins. Although efficient differentiation of pluripotent stem cells to hepatocytes has been achieved, the formation of proper polarisation in these cells has not been thoroughly investigated. In the present study, human embryonic stem cells (hESCs) and human mesench...

متن کامل

Differentiation of human embryonic stem cells into neurons

Human embryonic stem (ES) cells are undifferentiated pluripotent cells derived from the inner cell mass of blastocyst stage embryos. These unique cell lines have the potential to form virtually any cell type in the body and can be propagated in vitro indefinitely in an undifferentiated state. These cells are capable of forming embryoid bodies (EB) that contain cells from all three embryonic lin...

متن کامل

(hMSCs) were differentiated to hepatocyte-like cells and the derived hepatocytes were characterised for mature hepatocyte markers. The secretion of hepatic proteins, expression of hepatic genes and the functional hepatic polarisation of stem cell-derived hepatocytes, foetal hepatocytes and the HepG2

Adult hepatocytes are polarised with their apical and basolateral membranes separated from neighbouring cells by tight junction proteins. Although efficient differentiation of pluripotent stem cells to hepatocytes has been achieved, the formation of proper polarisation in these cells has not been thoroughly investigated. In the present study, human embryonic stem cells (hESCs) and human mesench...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Stem cells

دوره 26 5  شماره 

صفحات  -

تاریخ انتشار 2008