Can I plate IVT fresh hepatocyte suspensions and get these to attach?
IVT supplies fresh hepatocyte suspensions from human and other species without any guarantee that they will attach. However, we have talked to many of our customers who tell us that they have successfully plated our hepatocyte suspensions using the following technique: • Spin cells out of the suspension media at 50 g. • Resuspend the cells in IVT Hepatocyte Culture Media (to which you must first add 10% fetal bovine serum). • Perform a cell count; dilute to a final concentration of 0.7 million cells/mL in Hepatocyte Culture Media + 5% fetal bovine serum. • Add 0.5, 1.0, or 2.5 mL of this cell suspension to individual wells of a 24-, 12-, or 6-well cell culture plate, respectively. • Gently shake the plate in all directions (but not in a circular motion) to evenly distribute the cells in each well. • Place in a 37 degree Celsius, 5% CO2 incubator for 1–2 hours to allow cells to attach. • When cells have attached, wash off any unattached cells using pre-warmed Hepatocyte Culture Media +
