Cell-Bioink Mixing: Pipette Method

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cell bioink mixing pipette method

Overview

When bioprinting with a cell-laden bioink, it is important to thoroughly mix your biomaterial (or bioink) with your cells of choice in order to get a homogeneous cell distribution. This protocol for cell-bioink mixing using a pipette is a helpful guide for this key step in the bioprinting process.

Materials

Methods

  1. Calculate the number of cells needed. This will vary depending on the cell type and your final application; 
  2. Prepare a cell pellet (check out this protocol);
  3. If possible, pre-warm your sterile biomaterial to 37 °C to avoid temperature stress for the cells. Keep in mind that some materials should be kept at low temperatures to ensure good printability (e.g. collagen);
  4. With a pipette, add the appropriate volume of biomaterial to your cell pellet, keeping in mind how much material you need for your print as well as your desired cell concentration;
  5. Thoroughly mix cells with biomaterial by pipetting it up and down;
  6. Cap your Allevi 5 mL syringe barrel and flip the syringe so that the cap is facing down;
  7. Pipette your cell-laden biomaterial into the syringe barrel, dispensing material at the bottom of the syringe as uniformly as possible;
    • Note: be sure not to introduce bubbles to the system.
  8. Place your syringe stopper at the syringe opening;
  9. Flip the syringe barrel to have the cap facing up, ensuring to not let go of the stopper;
  10. Remove the syringe cap;
  11. Use the plunger to push material up until it reaches the syringe tip;
    • Note: be sure not to reconnect the plunger and the stopper.
  12. Tap the syringe to remove any bubbles that may have been trapped;
  13. Remove the plunger;
  14. Add your tip of choice for bioprinting;
  15. Bioprint!

Note: While calculating the necessary volume of your biomaterial, take into account dead-volume in the Luer-lock coupler and needle tips, which might affect the amount of material available for bioprinting. 

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