Tau-GFP genetically modified dissociated liver cells are isolated from Tau-GFP knock-in mice, 2-5 days after birth. These cells have a polygonal-shaped morphology after four day of culture.
Disaggregated mouse liver cells are isolated from mice, 2-5 days after birth and washed and digested in Kreb's Ringer Buffer (0.15 mM CalCl2 and 0.54 mg/ml of collagenase type I) for 40 min at 37°C. Cells are plated on collagen-coated plates in hepatocyte plating medium comprised of Dulbecco’s modified Eagle’s medium (DMEM) supplemented with bovine serum albumin (2 g/l), glucose (2 g/l), galactose (2 g/l), ornitine (0.1 g/l), proline (0.03 g/l), nicotinamide (0.61 g/l), Zncl2 (0.025 mg/l), ZnSO4:7H2O (0.75 mg/l), CuSO4:5H2O (0.2 mg/l), MnSO4 (0.025 mg/l), glutamine (5 mM), insulin (5 mg/l), human transferrin (5 mg/l), selenium (5 µg/l), dexamethasone (10-7 M), penicillin (100 mg/l), streptomycin (100 mg/l), and 10% calf serum. After 4 hours, the medium is changed to hepatocyte culturing medium that is comprised of serum-free hepatocyte plating medium supplemented with HGF (40 ng/ml) and EGF (20 ng/ml).
EMBRYONIC DEVELOPMENT & STEM CELL COMPENDIUM
