Human hepatocytes can be encapsulated in various bioinks for printing liver organoids or other geometries. Follow this protocol to encapsulate your hepatocytes. Please note that these hepatocytes cannot be passaged.
- Human Hepatocytes
- Hepatocyte Culture Medium
- Bioink of choice (Lifeink® 200, PhotoHA®, PhotoCol®, PureCol®, GelMA, Sterile GelMA, Glycosil®, Heprasil®, Allevi Liver dECM)
- Water/Bead Bath (37˚C)
- Biosafety Cabinet (BSC)
- 70% Ethanol
- Automatic Pipet Filler
- Serological Pipets
- Micropipette Tips
- Make sure you have properly thawed and counted your hepatocytes by following the Hepatocyte Thawing Protocol and the Counting Cells with a Hemocytometer protocols;
- Make sure to properly spray everything with 70% ethanol and wipe it with Kimwipes before placing anything in the BSC;
- Warm hepatocyte culture medium to 37˚C in a water/bead bath;
- Prepare your bioink of choice according to its preparation protocol. If possible, prepare your bioink with hepatocyte culture medium;
- The recommended cell density for hepatocyte encapsulation is 2.5 x 106 cells/mL, so add the appropriate amount of bioink to ensure you have that final concentration;
- Example: If you have counted a total of 5 million cells in your suspension, you will need a total of 2 mL of bioink. The formula is: total cells/cell concentration = amount of media to add to cell pellet (in mL).
- You may also follow this protocol, with an alternative cell-bioink mixing technique.
- Pipette it up and down to ensure homogenous cell distribution throughout the gel;
- Your cell-laden bioink is ready to be loaded into a syringe. To do so, follow this protocol.