Frequently Asked Questions - Hepatocytes
Can I repeatedly thaw and refreeze cryopreserved hepatocytes if I only need to remove a small aliquot of these products from a vial?
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.
Is it possible to "plate" cryopreserved human hepatocytes?
BioreclamationIVT 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 characterisation tables to find the lots that show excellent plating efficiency.Please contact our customer service team to enquire about purchasing these cells and they will happily liaise with BioreclamationIVT on your behalf.
Can I plate BioreclamationIVT fresh hepatocyte suspensions and get them to attach?
BioreclamationIVT 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 BioreclamationIVT 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 + 5% fetal bovine serum. 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 IVT Fresh Hepatocyte Suspensions].
BioreclamationIVT offers plated fresh hepatocytes in a variety of formats. Inquire with Customer Service on which formats are available.
Where can I find the fresh animal hepatocyte isolation calendar?
Go to the resource section of the website to view the fresh animal hepatocyte isolation schedule.
What seeding density should be used for a 96-well or 384-well plate?
Optimal seeding densities for human and most animal species are as follows:
96-well plate: 50,000 cells per well
384-well plate: 10,000 cells per well
Please note: we recommend mouse hepatocytes be seeded at lower densities.