Differentiation of human and mouse embryonic stem cells along a hepatocyte lineage
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Differentiation of human and mouse embryonic stem cells along a hepatocyte lineage. / Shirahashi, Hitoshi; Wu, Jian; Yamamoto, Naoki; Catana, Andreea; Wege, Henning; Wager, Brook; Okita, Kiwamu; Zern, Mark A.
in: CELL TRANSPLANT, Jahrgang 13, Nr. 3, 01.01.2004, S. 197-211.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Differentiation of human and mouse embryonic stem cells along a hepatocyte lineage
AU - Shirahashi, Hitoshi
AU - Wu, Jian
AU - Yamamoto, Naoki
AU - Catana, Andreea
AU - Wege, Henning
AU - Wager, Brook
AU - Okita, Kiwamu
AU - Zern, Mark A
PY - 2004/1/1
Y1 - 2004/1/1
N2 - Embryonic stem (ES) cells may differentiate along a hepatocyte lineage; however, currently there are no reports of culture conditions yielding high levels of hepatocyte-specific gene expression in these cells. We investigated culture conditions for differentiating ES cells into hepatocyte-like cells in vitro. Various combinations of culture media, growth and differentiation factors, and substratum precoatings were evaluated, and it was determined that a combination of Iscove's modified Dulbecco's medium with 20% fetal bovine serum, human insulin, dexamethasone. and collagen type I precoating was optimal for directing mouse ES cells along a hepatocyte lineage. Treatment of mouse ES cell with the optimal condition led to prealbumin gene expression 20% as high, and albumin synthesis 7% as high, as in mouse liver. The optimal culture condition also induced albumin gene expression in differentiated human ES cells 1% as high as in normal human hepatocytes as shown by Western blot analysis, and cells were positive for human albumin by immunocytochemistry. In addition, our optimal condition led to high levels of albumin gene expression in primary mouse hepatocytes after 35 days of culture, levels 10-fold higher than with other hepatocyte differentiation media. In conclusion, our optimal condition directed both mouse and human ES cells along a hepatocyte lineage. This represents the initial step in establishing cell lines that can be employed in cell-based therapeutics in humans and for toxicology and pharmacology studies.
AB - Embryonic stem (ES) cells may differentiate along a hepatocyte lineage; however, currently there are no reports of culture conditions yielding high levels of hepatocyte-specific gene expression in these cells. We investigated culture conditions for differentiating ES cells into hepatocyte-like cells in vitro. Various combinations of culture media, growth and differentiation factors, and substratum precoatings were evaluated, and it was determined that a combination of Iscove's modified Dulbecco's medium with 20% fetal bovine serum, human insulin, dexamethasone. and collagen type I precoating was optimal for directing mouse ES cells along a hepatocyte lineage. Treatment of mouse ES cell with the optimal condition led to prealbumin gene expression 20% as high, and albumin synthesis 7% as high, as in mouse liver. The optimal culture condition also induced albumin gene expression in differentiated human ES cells 1% as high as in normal human hepatocytes as shown by Western blot analysis, and cells were positive for human albumin by immunocytochemistry. In addition, our optimal condition led to high levels of albumin gene expression in primary mouse hepatocytes after 35 days of culture, levels 10-fold higher than with other hepatocyte differentiation media. In conclusion, our optimal condition directed both mouse and human ES cells along a hepatocyte lineage. This represents the initial step in establishing cell lines that can be employed in cell-based therapeutics in humans and for toxicology and pharmacology studies.
KW - Albumins
KW - Animals
KW - Blotting, Western
KW - Cell Culture Techniques
KW - Cell Differentiation
KW - Cell Lineage
KW - Cell Transplantation
KW - Culture Media
KW - Dexamethasone
KW - Embryo, Mammalian
KW - Hepatocytes
KW - Humans
KW - Immunohistochemistry
KW - Insulin
KW - Mice
KW - Models, Biological
KW - Species Specificity
KW - Stem Cells
KW - Time Factors
KW - Urea
M3 - SCORING: Journal article
C2 - 15191158
VL - 13
SP - 197
EP - 211
JO - CELL TRANSPLANT
JF - CELL TRANSPLANT
SN - 0963-6897
IS - 3
ER -