hepatocytes derived stem cell - hepatocytes derived stem cell (Jan/23/2005 )

Does anyone known how to differentiate stem cell to hepatocytes. Thank you.

We cultured stem cells in the base medium: DMEM/Hams’ F12 (1:1) containing 1% ITS culture supplement (BD Bioscience), 0.1 nM MEM nonessential amino acids solution (Invitrogen), 5 mM MEM sodium pyruvate (Invitrogen), 10 mM nicotinamide (Sigma) and 0.2 mM L-ascorbic acid-2-phosphate (Sigma).

The base medium was further supplemented with the following differentiation factors:
10 ng/mL acidic fibroblast growth factor (Epoch Biolabs);
20 ng/mL hepatocyte growth factor (Epoch Biolabs);
1 uM 5-Aza-2-deoxycytidine (5AC) (Sigma);
1 uM Dex (Sigma);
cell incubation for 7-10 days.

The expression of hepatic proteins, such as alpha-fetoprotein (AFP), albumin (ALB), cytokeratin 8 (CK8), cytokeratin 18 (CK18), in cytoplasm was analyzed by immunocytochemistry (ICC).

ps. We found that aFGF and HGF from Epoch Biolabs gave us the most consistent results over other vendors.

There has been many a paper about the differentiation,characterization of stem cell derived hepatocyte or hepatocyte-like cells ,just google for them, you'll find ther're different strateges

Carlos Semino et al at MIT showed they could take a line of hepatocyte/liver progenitor cells (James Sherley's Lig8 line) and differentiate them into mature hepatocytes in PuraMatrix synthetic nanofiber scaffolds. This defined 3D cell culture matrix enabled defined reproducible cell culture microenvironments for controlled stem cell differentiation, and superior to collagen sandwich gels and Matrigel.

Hope it helps!

Semino, C.E., Merok, J., Crane, G., Panagiotakos, G., Zhang, S. Functional differentiation of hepatocyte-like spheroid structures from putative liver progenitor cells in three-dimensional peptide scaffolds. Differentiation 71:262–270 (2003).

Also see http://www.puramatrix.com/pubs