P431 Aims: In order to easily reproduce the results of human islets isolations, we developed and evaluated a simplified digestion technique using a polymethylpenten chamber (PMPC) and compared this method with the automated islet isolation technique using a stainless steel chamber (SSC), described by Ricordi et al. (Diabetes, 1989) Methods: Twenty human pancreases obtained from cadaveric donors and preserved in UW solution were digested with either SSC (Group 1, n=5) or PMPC (Group 2, n=15). There was no significant difference between the two groups in terms of donors characteristics (age, CIT, WIT, BMI, amylase level). The pancreases digested with SSC were processed exactly following Ricordi’s method. Digestion with PMPC was performed with Liberase HI at 37°C without any circulation system. The digested tissue was collected after filtration through a 1000μm mesh screen and passed at 4°C on a column containing 6-mm glass beads and a 500μm mesh screen. Results: PMP chamber maintained a stable temperature at 37°C during the entire pancreas digestion. No significant difference was observed in term of pancreas digested tissue (>70%) between 2 groups. The islets equivalent yield by gram of digested tissue (IEQ/gram) was similar with both methods (5520 IEQ/gr for PMPC vs. 4679 IEQ/gr for SSC, p>0.05). A higher percentage of large islets (>100μm) was obtained with PMP chamber (p<0.05). The final volume obtained after pancreas digestion with PMPC (13±5 ml) was significantly reduced (4 fold) in comparison with the tissue volume obtained with SSC digestion (51±14ml, p<0.005).This small volume facilitated the purification step without Cobe 2991. Islet recovery (IEQ after digestion/IEQ after purification) with a simple discontinuous gradient (Group 2, n=15) was 63% and islets viability/purity reached 84±8% and 73±20%, respectively. The final cellular volume obtained with this technique, reached a mean of 880μl. Immunohistochemistry evidenced a similar (p>0.05) proportion of beta cells prior (56±10%)(n=6) and after (61±10%)(n=9) isolation. By transmission electron microscopy, the good preservation of the isolated islets containing many granulated cells was established. Islet function, assessed by in vitro systems (Glucose 1-15mM) revealed a stimulation index at 13.1±7.1. Three highly purified islets transplantation were carried out by intra-portal injection without any increase of portal pressure or changes of liver enzymes. In two cases, a significant decrease of daily insulin requirement (-40%) and no hypoglycemia (increase quality of life), after a single islets injection (<5,000 IEQ/kg) was observed at hospital discharge (follow-up: 12 months). Conclusions: Digestion technique of human pancreas with PMPC provided a simple method of pancreatic islets isolation, an equal yield of islets than previously described digestion methods as well as a preserved morphological integrity, viability and functionality. Last but not least, this technique might be carried out by only two persons and is significantly less expensive since all material are “one use” which is overwhelming for good medical practice.