Splitting Spheroids for Sub-culturing and Shipping: A Procedure for Propagating and Transferring Spheroids from Small Bowel Neuroendocrine Tumor

Mechanically break the ECM with a P1000 pipette and transfer it to a sterile 1.5-millimeter tube. Centrifuge the tube at 1,000 x g at 4 degrees Celsius. Then, remove all supernatant and place the tube on ice. Add two to four times the volume of new ECM to the pellet and mix the contents of the tube by pipetting up and down 10 times, making sure to avoid introducing air bubbles.

Transfer 5 to 20 microliters of the ECM in SBNET mixture to a new 96-well plate and allow the ECM to solidify. Then, cover the solidified ECM with new SBNET medium and put the plate in the incubator. If shipping SBNET spheroids to another lab, transfer the spheroids with the new ECM to a T25 flask and allow the ECM to solidify. Fill the flask with SBNET spheroid medium, screw the cap on tightly, and prepare the shipping package.