Protocol: P3D Scaffold Cell Recovery by Trypsination
This protocol explains how to retrieve cells that have been cultured on the Ossiform P3D Scaffolds. Cell recovery from the P3D Scaffolds can be performed using standard protocols with minor modifications.
Trypsin, accutase, or other appropriate enzymes.
Ilex Life Sciences offers recombinant trypsin free of animal and human sources:
- Bovine Trypsin, recombinant, expressed in corn, lyophilized powder
- Porcine Trypsin, E. coli recombinant, lyophilized powder
Notes before starting and general advice on material handling
- All handling of the P3D Scaffolds products should be performed using gloves, according to the standard aseptic methods.
- In general, 3D cultures require longer incubation times and more rigorous shaking than 2D cultures.
Cell recovery from the P3D scaffolds can be performed using standard protocols with small modifications.
- Make sure that the enzymatic agent completely covers the scaffolds (minimum 300 uL pr well for 24-well plates).
- Incubate for 3-5 minutes.
- Stop the trypsinization by adding media and transfer liquid and scaffold to a clean 50 mL Falcon tube.
- Centrifuge for 5 minutes at 1200-4000 rpms to retrieve cells “dislodged” inside the scaffold as well.
- Remove scaffold and resuspend cell pellet by pipetting up and down.
The products are “For Research Use Only (RUO)” and should not be used for clinical, diagnostic, or therapeutic purposes. Ossiform and its distributor, Ilex Life Sciences LLC, make no other warranties, expressed or implied, including the implied warranty of merchantability and the implied warranty of fitness for particular purpose.
This protocol has been adapted from the protocol published by Ossiform. Ilex Life Sciences is an authorized distributor of Ossiform P3D Scaffolds research product line.