Human Pancreatic Islet Isolation (Research Paper Sample)

Human Pancreatic Islet Isolation Name: Institution: Human Pancreatic Islet Isolation This process is a very critical procedure that is time sensitive hence requires a lot of team effort to be completed successfully. The head surgeon leads this entire process and it begins by ensuring that each of the equipment that may be used in the isolation is working and in good condition. This means that all working areas have to be sanitized while machines such as the centrifuge has to be tested to ensure it is fully functioning. The areas that need to be properly set up include the surgical tables, perfusion units, cannulation table, digestion circuit, slush machine and the COBE purification machine. Additionally, sampling tubes that have been prefilled with wash, bacteriology tubes and assessment tubes also have to be prepared beforehand. To begin the process, the purification tubes are set in such a way that the first one is empty and marked at the 150ml mark while the other 11 tubes are filled with doses of M199 mixed with human albumin (wash solution) up to the 200ml mark (Shaw & Shapiro, 2007, p.172). The last step involves filling the 50ml conical tube with wash solution after which the process of preparing the pancreas can begin. First of all, one of the operators has to remove the pancreas from its package using a sterile method after which the surgeon will transfer it to the decontamination table. The surgeon is then able to fill up a 60ml syringe with the original pancreas preservation solution to prepare the bacteriology bottles for later use. In addition to that, the surgeon also carries out a visual inspection of the pancreas to ensure its suitability for the procedure before asking on of the operators to prepare collagenase. It is at this point that the surgeon can bring the decontamination solutions to the table and pour them into the proper containers (Stratta & Hakim, 2010, p.238). These solutions include Betadine, Fungizone/Cefazolin and HBSS. The surgeon will now be able to decontaminate the pancreas using all the above solutions one at a time after which it is placed in a sterile jar where its weight is measured then placed in a cannulation pan with preservation solution. The surgeon will next incise the main duct and expose it after which each half is cannulated. The pancreas is then perfused by placing it in the perfusion unit and pouring an enzyme solution on the unit for about 10 minutes whereby 5 minutes will be used to perfuse it at 80mmHg and the other 5 minutes at 180mmHg. This perfused pancreas is then moved to the cannulation pan where the surgeon will trim off fatty tissue and cut it into 10-12 pieces that will be transferred to the Ricordi Chamber to begin digestion. Once in the chamber, 2.5 ml vial of Pulmozyme is added after which it is closed and the system is filled up. The solution has to be pumped for 5 minutes after which the chamber can be rocked gently to disperse the warm solution evenly while m...