Although the transplantation of alginate-poly-L-lysine-alginate encapsulated islets of Langerhans usually is successful, graft survival is still limited. Molecular analysis by RT-PCR of the encapsulated islets may provide insight into the mechanisms that affect islets during graft failure. However, RT-PCR on encapsulated islets is not possible because the poly-L-lysine of the capsule interferes with both cDNA synthesis and PCR amplification. We applied a method that mechanically removes the microcapsules from the islets after a short trypsin-EDTA treatment (decapsulation), thereby enabling RT-PCR analysis. The results of this study show that the decapsulation procedure does not affect islet vitality and has only minor effects on islet function and morphology. The decapsulation does not affect GAPDH, beta-actin, Bcl-2, or Bax gene expression. This method is an improvement over the time-consuming manual dissection of microcapsules because it allows for the rapid and relatively harmless removal of capsules on a larger scale. Decapsulation offers the possibility of applying RT-PCR, as well as other methods, which cannot be performed on encapsulated islets.
Islet transplantation has proven to be a potential cure for insulin-dependent diabetic patients (Shapiro et al. 2000). Islet isolation is crucial for successful islet transplantation. Islet mass, islet quality and islet purity are determined by islet isolation ef cacy, which depends on many factors, including organ procurement, collagenase digestion and puri cation method (Williams et al. 1995, Vos-Scheperkeuter et al. 1997, Morrison et al. 2002. Apart from differences in islet yield among species (van Suylichem et al. 1995, Wieczorek et al. 1998, variability in islet yield within one species is also found (van Deijnen et al. 1992). These variations can be attributed to organ quality (van der Burg et al. 1994b), batch-to-batch variation in collagenase preparation (Wolters et al. 1990(Wolters et al. , 1992, and donor age
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