Pancreas Dissociation
Pancreas dissociation and harvest SOP: Pancreas dissociation and tissue harvest
Pancreas Dissociation and harvest
Once heated the enzymes initiate enzymatic digestion of the pancreatic macrostructure. To accomplish this, as seen in the Enzyme Perfusion video on the previous page, the perfused organ is cut into smaller pieces and placed in an automated dissociation device with warm, recirculating solution. Marbles help to mechanically dissociate the tissue and a series of three filters keep larger, undissociated tissue inside the device.
Monitoring the digest is an important step and requires experienced personnel to determine when to harvest. Samples are dithizone stained during the recirculation phase.
Once enough tissue of good quality passes through the filters, the system is opened for harvesting. Collected tissue is immediately cooled in conicals and the enzymes are inactivated by serum to halt further, unwanted damage to the tissue. This procedure continues for about 40 minutes and uses about 7-8 liters of solution.