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Effectiveness of hydroxyethyl starch (HES) on purification of pancreatic islets
Takashi Kenmochi1, Takehide Asano, Kazuhiko Jingu
1Department of Academic Surgery (M9), Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba City, Chiba 260-8670, Japan. kenmochi@snh.hosp.go.jp
Background:
The isolation of large-scale and high-quality islets from the pancreas is essential for a successful islet transplantation. We developed a hydroxyethyl starch (HES)-Collins solution as the purification medium and evaluated its usefulness on islet isolation from the pancreas of beagle dogs.
Materials And Methods:
The pancreas of beagle dog was digested using our original 2-step automated technique. Islets were purified by discontinuous purification method on HES-Collins solution (group 1, n = 10) or Euro-Ficoll solution (group 2, n = 16) with a COBE2991 cell processor. Islet yield and purity, changes in islet number during 3-day cultures, static incubation, perifusion study, and insulin content were examined. In addition, the islets were autotransplanted into the liver via the portal vein.
Results:
Although no significant differences were detected, yield and purity were higher in group 1. After 3 days of culture, the islet number decreased less in group 1. Both under static incubation and in the perifusion study, stimulation indices were 4.50 +/- 1.82 and 6.02 +/- 1.05, which were significantly higher than the 2.18 +/- 0.67 and 3.32 +/- 1.46 observed in group 2. Also, insulin content of the islets was significantly higher in group 1 than in group 2. Fasting blood glucose levels were maintained at values below 100 mg/dl for 100 days in group 1 and around 200 mg/dl in group 2.
Conclusions:
The use of HES-Collins solution was associated with an improvement of islet yield and purity. Also, islet viability and function were preserved longer during culture. Accordingly, islet purification using HES-Collins solution might be recommended for clinical islet transplantation.