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.