DataSheet_1_Optimal temperature for the long-term culture of adult porcine islets for xenotransplantation.pdf

Porcine islet xenotransplantation represents a promising therapy for severe diabetes mellitus. Long-term culture of porcine islets is a crucial challenge to permit the on-demand provision of islets. We aimed to identify the optimal temperature for the long-term culture of adult porcine islets for xenotransplantation. We evaluated the factors potentially influencing successful 28-day culture of islets at 24°C and 37°C, and found that culture at 37°C contributed to the stability of the morphology of the islets, the proliferation of islet cells, and the recovery of endocrine function, indicated by the expression of genes involved in pancreatic development, hormone production, and glucose-stimulated insulin secretion. These advantages may be provided by islet-derived CD146-positive stellate cells. The efficacy of xenotransplantation using islets cultured for a long time at 37°C was similar to that of overnight-cultured islets. In conclusion, 37°C might be a suitable temperature for the long-term culture of porcine islets, but further modifications will be required for successful xenotransplantation in a clinical setting.</p

DataSheet_3_Optimal temperature for the long-term culture of adult porcine islets for xenotransplantation.pdf

Porcine islet xenotransplantation represents a promising therapy for severe diabetes mellitus. Long-term culture of porcine islets is a crucial challenge to permit the on-demand provision of islets. We aimed to identify the optimal temperature for the long-term culture of adult porcine islets for xenotransplantation. We evaluated the factors potentially influencing successful 28-day culture of islets at 24°C and 37°C, and found that culture at 37°C contributed to the stability of the morphology of the islets, the proliferation of islet cells, and the recovery of endocrine function, indicated by the expression of genes involved in pancreatic development, hormone production, and glucose-stimulated insulin secretion. These advantages may be provided by islet-derived CD146-positive stellate cells. The efficacy of xenotransplantation using islets cultured for a long time at 37°C was similar to that of overnight-cultured islets. In conclusion, 37°C might be a suitable temperature for the long-term culture of porcine islets, but further modifications will be required for successful xenotransplantation in a clinical setting.</p

DataSheet_7_Optimal temperature for the long-term culture of adult porcine islets for xenotransplantation.docx

Porcine islet xenotransplantation represents a promising therapy for severe diabetes mellitus. Long-term culture of porcine islets is a crucial challenge to permit the on-demand provision of islets. We aimed to identify the optimal temperature for the long-term culture of adult porcine islets for xenotransplantation. We evaluated the factors potentially influencing successful 28-day culture of islets at 24°C and 37°C, and found that culture at 37°C contributed to the stability of the morphology of the islets, the proliferation of islet cells, and the recovery of endocrine function, indicated by the expression of genes involved in pancreatic development, hormone production, and glucose-stimulated insulin secretion. These advantages may be provided by islet-derived CD146-positive stellate cells. The efficacy of xenotransplantation using islets cultured for a long time at 37°C was similar to that of overnight-cultured islets. In conclusion, 37°C might be a suitable temperature for the long-term culture of porcine islets, but further modifications will be required for successful xenotransplantation in a clinical setting.</p

DataSheet_6_Optimal temperature for the long-term culture of adult porcine islets for xenotransplantation.pdf

Porcine islet xenotransplantation represents a promising therapy for severe diabetes mellitus. Long-term culture of porcine islets is a crucial challenge to permit the on-demand provision of islets. We aimed to identify the optimal temperature for the long-term culture of adult porcine islets for xenotransplantation. We evaluated the factors potentially influencing successful 28-day culture of islets at 24°C and 37°C, and found that culture at 37°C contributed to the stability of the morphology of the islets, the proliferation of islet cells, and the recovery of endocrine function, indicated by the expression of genes involved in pancreatic development, hormone production, and glucose-stimulated insulin secretion. These advantages may be provided by islet-derived CD146-positive stellate cells. The efficacy of xenotransplantation using islets cultured for a long time at 37°C was similar to that of overnight-cultured islets. In conclusion, 37°C might be a suitable temperature for the long-term culture of porcine islets, but further modifications will be required for successful xenotransplantation in a clinical setting.</p

DataSheet_4_Optimal temperature for the long-term culture of adult porcine islets for xenotransplantation.pdf

Porcine islet xenotransplantation represents a promising therapy for severe diabetes mellitus. Long-term culture of porcine islets is a crucial challenge to permit the on-demand provision of islets. We aimed to identify the optimal temperature for the long-term culture of adult porcine islets for xenotransplantation. We evaluated the factors potentially influencing successful 28-day culture of islets at 24°C and 37°C, and found that culture at 37°C contributed to the stability of the morphology of the islets, the proliferation of islet cells, and the recovery of endocrine function, indicated by the expression of genes involved in pancreatic development, hormone production, and glucose-stimulated insulin secretion. These advantages may be provided by islet-derived CD146-positive stellate cells. The efficacy of xenotransplantation using islets cultured for a long time at 37°C was similar to that of overnight-cultured islets. In conclusion, 37°C might be a suitable temperature for the long-term culture of porcine islets, but further modifications will be required for successful xenotransplantation in a clinical setting.</p

DataSheet_5_Optimal temperature for the long-term culture of adult porcine islets for xenotransplantation.pdf

Porcine islet xenotransplantation represents a promising therapy for severe diabetes mellitus. Long-term culture of porcine islets is a crucial challenge to permit the on-demand provision of islets. We aimed to identify the optimal temperature for the long-term culture of adult porcine islets for xenotransplantation. We evaluated the factors potentially influencing successful 28-day culture of islets at 24°C and 37°C, and found that culture at 37°C contributed to the stability of the morphology of the islets, the proliferation of islet cells, and the recovery of endocrine function, indicated by the expression of genes involved in pancreatic development, hormone production, and glucose-stimulated insulin secretion. These advantages may be provided by islet-derived CD146-positive stellate cells. The efficacy of xenotransplantation using islets cultured for a long time at 37°C was similar to that of overnight-cultured islets. In conclusion, 37°C might be a suitable temperature for the long-term culture of porcine islets, but further modifications will be required for successful xenotransplantation in a clinical setting.</p

DataSheet_2_Optimal temperature for the long-term culture of adult porcine islets for xenotransplantation.pdf

Porcine islet xenotransplantation represents a promising therapy for severe diabetes mellitus. Long-term culture of porcine islets is a crucial challenge to permit the on-demand provision of islets. We aimed to identify the optimal temperature for the long-term culture of adult porcine islets for xenotransplantation. We evaluated the factors potentially influencing successful 28-day culture of islets at 24°C and 37°C, and found that culture at 37°C contributed to the stability of the morphology of the islets, the proliferation of islet cells, and the recovery of endocrine function, indicated by the expression of genes involved in pancreatic development, hormone production, and glucose-stimulated insulin secretion. These advantages may be provided by islet-derived CD146-positive stellate cells. The efficacy of xenotransplantation using islets cultured for a long time at 37°C was similar to that of overnight-cultured islets. In conclusion, 37°C might be a suitable temperature for the long-term culture of porcine islets, but further modifications will be required for successful xenotransplantation in a clinical setting.</p

Condition of PHLDA3-deficient islets at 24 hs after isolation.

<p><b>A.</b> Islets from PHLDA3 KO and WT groups were stained by SYTO Green 11 (for viable cells) and ethidium bromide (for dead cells, indicated by arrow). More viable cells and fewer dead cells were seen in KO group than the WT group. The size of the scale bar is 100 μm. <b>B.</b> Quantification of islet viability showed significantly higher viability for PHLDA3-deficient (= KO: n = 38 islets) than wild-type islets (n = 31 islets). <b>C.</b> Volumes of released insulin following stimulation in low (3.3mM) and high (16.5mM) glucose solutions. Insulin release dependent on glucose concentration was seen in both the KO and WT groups, and the levels between the two groups were similar (n = 15 islets in both groups). For statistical analysis, Mann–Whitney U test was used for viability and two-way repeated measures ANOVA for insulin—releasing function.</p

Metabolic condition of diabetic mice after intrahepatic islet transplantation with PHLDA3-deficient or wild-type islets.

<p>Blood glucose (<b>A</b>) and C-peptide (<b>B</b>) levels were measured after transplantation of PHLDA3-deficient or wild-type islets, and the difference between the KO (n = 17) and WT (n = 14) groups was statistically assessed by two-way repeated measures ANOVA. Levels of both were significantly improved in KO group in comparison with WT group. Blood glucose levels gradually decreased in both groups, but were significantly lower in the KO group compared to the WT group by postoperative day (POD) 28 (357.3 ± 35.7 mg/dL in KO group vs. 470.4 ± 35.4 mg/dL in WT group at POD 28, p = 0.04; 286.8 ± 33.3 mg/dL vs. 428.6 ± 42.4 mg/dL at POD 56, p = 0.02; <b>A</b>). Change in blood glucose levels after glucose stimulation (<b>C</b>) and AUC of the blood glucose levels after glucose stimulation (<b>D</b>) at POD 56 also revealed that the metabolic condition of the mice transplanted with PHLDA3-deficient islets was improved. The times of experiments were eight. For statistical analyses, the two-way repeated measures ANOVA for change of blood glucose level and Mann–Whitney U test for AUC were used.</p

Size and number of engrafted PHLDA3-deficient islets.

<p><b>A.</b> Engrafted islets at POD 56 and 180. The size was almost the same between the KO and WT groups at POD 56 (upper and middle lanes). Hyperplasia of PHLDA3-deficient islets was not detected at POD 180, but the size tended to be wider than that at POD 56 (bottom vs top panels). The size of the scale bar was 100 μm. <b>B</b> and <b>C</b>. Quantification of islet areas. There was no difference between the KO (n = 29 islets) and WT (n = 8 islets) groups at POD 56 (<b>B</b>). There was also no difference in sizes between POD 56 (n = 29 islets) and 180 (n = 7 islets) in the KO group, but the islet area at 180 tended to be wider than that at POD 56 (<b>C</b>). <b>D</b>. Numbers of engrafted islets at POD 56. More engrafted islets were seen in the KO group (n = 17) than the WT group (n = 14). Mann–Whitney U test was used for statistical assessment.</p