raffinose and iodixanol

Sufficient and favorable biological functions of islets are major problems hindering xenotransplantation. The aim of the present study was to evaluate the effects on harvesting, purity, viability, and function of using improved Visipaque (iodixanol) and Ficoll-400 for adult porcine islet purification.. Twelve adult porcine pancreata were randomly divided into an Iodixanol-University of Wisconsin (UW) group and a Ficoll-400-UW group according to the purification method. Porcine pancreata were isolated by collagenase digestion. After isolation and purification, the islet yield and purity were evaluated by dithizone staining, and islet function assessed by in vitro insulin release assays and in vivo islet xenotransplantation.. There were no marked differences in the islet yield before purification (5254.67 +/- 189.44 IEQ/g vs 5092.67 +/- 178.94 IEQ/g, P > .05). After purification, there were significantly more islets harvested in Iodixanol-UW group than in the Ficoll-400-UW group: 4222.00 +/- 228.84 IEQ/g vs 3036.83 +/- 79.60 IEQ/g (P < .05). Islets from the two groups showed satisfactory insulin secretory ability. There were no significant differences in islet survival times between the two groups in diabetic rats: 8.2 +/- 1.619 days vs 6.9 +/- 1.197 days (P > .05).. The improved iodixanol-UW density gradient method was superior to Ficoll-400 method to improve the number, viability, and insulin secret of purified adult porcine islets although the benefits did not improve in vivo survival.

Topics: Adenosine; Allopurinol; Animals; Blood Glucose; Cell Count; Cell Separation; Cell Survival; Contrast Media; Diabetes Mellitus, Experimental; Ficoll; Glutathione; Insulin; Insulin Secretion; Islets of Langerhans; Islets of Langerhans Transplantation; Organ Preservation Solutions; Raffinose; Rats; Rats, Sprague-Dawley; Swine; Transplantation, Heterologous; Triiodobenzoic Acids

Primary cells of renal proximal tubule epithelium (S1 segment) of human kidney (HRPTE cells) up-regulate aquaporin-1 (AQP-1) expression in response to hyperosmolarity. NaCl and D(+)-raffinose increased (2-2.5 fold) AQP-1 expression when medium osmolarity was 400 and 500 mOsm/kg.H2O. Urea did not have this effect. Unlike our previous findings with mIMCD-3 cells, vasopressin (10(-8)M) did not affect AQP-1 expression in HRPTE cells in isosmolar or NaCl-enriched hyperosmolar conditions. Furthermore, HRPTE cells increased (3-4 fold) AQP-1 expression when exposed to hyperosmolar Reno-60 and Hypaque-76 (diatrizoates, ionic) contrast agents at 400 and 500 mOsm/kg.H2O. Isosmolar (290 mOsm/kg H2O) Visipaque (iodixanol, non-ionic) at 10% (v/v) concentrations also increased AQP-1 expression, and 25% v/v of Visipaque rendered morphological alterations of HRPTE cells and a 3-fold increase in AQP-1 expression after 24h exposure. Finally, semi-quantitative RT-PCR of HRPTE cells subjected to various isosmolar or hyperosmolar conditions demonstrated up-regulation of AQP-1 mRNA and protein levels. Our results suggest AQP-1 up-regulation in HRPTE cells exposed to environmental stresses such as hyperosmolarity and high doses of isosmolar contrast agents.

Topics: Aquaporins; Blotting, Western; Cell Size; Cells, Cultured; Contrast Media; Diatrizoate; Diatrizoate Meglumine; Drug Combinations; Epithelial Cells; Humans; Iohexol; Kidney Tubules, Proximal; Osmolar Concentration; Raffinose; Reverse Transcriptase Polymerase Chain Reaction; Sodium Chloride; Time Factors; Triiodobenzoic Acids; Up-Regulation; Urea

Topics: Adenosine; Allopurinol; Animals; Cell Separation; Centrifugation, Density Gradient; Glutathione; Insulin; Islets of Langerhans; Islets of Langerhans Transplantation; Organ Preservation Solutions; Raffinose; Swine; Triiodobenzoic Acids