FAQ - Hepatocytes
No. We have found that repeated freezing and thawing causes extremely low recovery of viable hepatocytes. That is why we pack our cryopreserved hepatocytes in a volume that is convenient for most single-use applications.
BioIVT offers cryoplateble human hepatocytes. These cells act like fresh human hepatocytes in that they plate and attach for 5 days and respond well to inducers. Please refer to the characterization tables to find the lots that show excellent plating efficiency. Contact BioIVT to inquire about purchasing these cells.
BioIVT 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 BioIVT INVITROGRO™ CP medium, perform a cell count and dilute to a final concentration of 0.7 million cells/mL. 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 north-south and east-west pattern to evenly distribute the cells in each well. Place in a 37 degree Celsius, 5% CO2 incubator for 3-4 hours to allow cells to attach. When cells have attached, wash off any unattached cells using pre-warmed INVITROGRO CP medium. Incubate overnight. The cells are now ready to be used in your experiments.
[Note: This is just one of many possible methods for attempting to plate the BioIVT Fresh Hepatocyte Suspensions].
BioIVT offers plated fresh hepatocytes in a variety of formats. Inquire with Customer Service on which formats are available.
Got to the resource section of the web site to view the fresh animal hepatocyte isolation schedule.
Optimal seeding densities for human and most animal species are as follows:
Please note: Mouse hepatocytes are larger than that of other species and require half the seeding density. (for example, 0.35 x 106 viable cells/mL instead of 0.70 x 106 viable cells/mL)
For more top tips for hepatocyte handling check out our blog.