A simple method for the release of islets by controlled collagenase digestion of the human pancreas.

摘要: A simple technique for the controlled collagenase digestion of human pancreas is described. The distended with collagenase, and a biopsy taken divided into 5 pieces that are placed in Universals containing minimal essential medium dithizone at 39 degrees C. itself incubated MEM Starting min intervals thereafter, Universal removed from water bath, shaken 30 sec, contents examined by microscopy. As soon as free cleaved islets seen, one compartment kidney-bowl half 1-mm mesh. gently teased apart fluid digest empty bowl aspirated passed through 500-micron mesh ice-cold 20% newborn calf serum. This process repeated until has ceased. Using this on 20 consecutive pancreata, median wt. (range) 53.9 (45.2-72.9) g, we have counted 131,672 (43,516-400,000) digest, equivalent to 2394 (715-8000) islets/g pancreas. volume islet tissue was 299 (26-1341) mm3 5.81 (0.36-26.81) mm3/g In conclusion, found be an effective method