Abstract
Robust generation of hepatocyte-like cells from human embryonic stem cell populations
Highlights
Despite progress in modelling human drug toxicity, many compounds fail during clinical trials due to unpredicted side effects
In order to characterise the stem cell status of the H9 hESCs used in the study we studied a number of parameters
The cells exhibited hESC morphology, small, tightly packed cells growing in defined colonies (Figure 1A) and expressed the pluripotent stem cell gene markers, Oct-3/4 and Nanog (Figure 1B)
Summary
All steps to be carried out in a tissue culture hood under aseptic conditions. 1. Prepare 10% BSA solution in PBS and filter through a 0.22 μm filter. 6. Aliquot the hbFGF in sterile eppendorfs and store at -20°C. 2. Preparation of Human Activin A Stock Solution 1. 3. Filter the Activin A solution and aliquot in sterile eppendorfs, store at -20°C. 4. Preparation of Human HGF Stock Solution (1000X) 1. 2. Filter the HGF solution and aliquot in sterile eppendorfs, store at -20°C. 2. Filter the OSM solution and aliquot in sterile eppendorfs, store at -20°C. Thaw the 10 mL stock bottle of Matrigel overnight at 4°C on ice and add 10 mL of KO-DMEM. 3. Add 5 mL of cold KO-DMEM to the matrigel, mix well with a pipette. 7. Plates or flasks which have been coated with matrigel can be stored at 4°C for up to 1 week. Prior to use aspirate the matrigel and add the cell suspension to the well or flask
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