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In this study, nontreated, type IV collagen, Lipidure, and ultralow attachment dishes were used to culture a murine \u03b2\u2010cell line, MIN\u20106. The formation and biological performances of pseudoislets were investigated. Results showed that \u03b2\u2010cells formed loose and irregular aggregates on nontreated dishes. Oppositely, pseudoislets formed on other three substrates. Most pseudoislets on Lipidure and type IV collagen dishes had a diameter between 100\u2013150 \u03bcm with high survival rate, while large pseudoislets (&gt;250 \u03bcm) with seriously central necrosis were found on ultralow attachment dishes. Western blot analysis revealed that pseudoislets had relatively higher connexin 36 protein productions relative to single cells. The glucose\u2010stimulated insulin secretion test showed pseudoislets on type IV collagen have high stimulation index. Monolayers from TCPS dishes and pseudoislets from type IV collagen or Lipidure dishes were further transplanted into diabetic mice. Animals received both single cells and pseudoislets had decreasing blood glucose level and regained body weight. Histologic examination revealed that all implants successfully engrafted with positive insulin staining. Interestingly, the area under curve for the intraperitoneal glucose tolerance test showed pseudoislets had superior glucose disappearance rate. This study reveals that isolated islets or insulin\u2010producing cells can be cultured on type IV collagen or Lipidure dishes to improve\/maintain integrity prior to transplantation. \u00a9 2013 Wiley Periodicals, Inc. 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