Published online Jan 28, 2005. doi: 10.3748/wjg.v11.i4.545
Revised: April 15, 2004
Accepted: September 1, 2004
Published online: January 28, 2005
AIM: To study the core cell damage in isolated islets of Langerhans and its prevention by low temperature preconditioning (26 °C).
METHODS: Islets were cultured at 37 °C for 7-14 d after isolation, and then at 26 °C for 2, 4 and 7 d before additional culture at 37 °C for another 7 d. Core cell damage in the isolated islets was monitored by video-microscopy and analyzed quantitatively by use of a computer-assisted image analysis system. The analysis included daily measurement of the diameter and the area of the isolated islets and the area of the core cell damage that developed in those islets over time during culture. Histology and TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay were used to characterize the cell damage and to monitor islet function.
RESULTS: Microscopic analysis showed that during the 7 to 14 d of culture at 37 °C, core cell damage occurred in the larger islets with diameters >200 μm, which included both necrotic and apoptotic cell death. Low temperature (26 °C) culture could prevent core cell damage of isolated islets. The 7-d culture procedure at 26 °C could inhibit most of the core cell (excluding diameters>300 μm) damages when the islets were re-warmed at 37 °C.
CONCLUSION: Our results indicate that core cell damage within isolated islets of Langerhans correlates with the size of islets. Low temperature (26 °C) culture can prevent core cell damage in isolated islets, and successfully precondition these islets for incubation at 37 °C. These novel findings may help to understand the pathophysiology of early loss of islet tissue after transplantation, and may provide a new strategy to improve graft function in the clinical setting of islet transplantation.