Reduced Porcine Islet Isolation Yield in the Presence of Hyperemic Islets

Abstract

When studying histological characteristics of porcine pancreata in relation to islet isolation, a remarkably high number of hyperemic islets (HIs) was encountered. The abnormalities observed in these HIs ranged from a single dilated vessel to hemorrhages extending into the surrounding exocrine tissue. The aim of the present study was to compare pancreata with and without HI on islet isolation outcomes. This study involved a histological examination of 143 purebred (74 juvenile and 69 adult) and 47 crossbred (only juvenile) porcine pancreata. Islet isolation was performed in 48 purebred adult pigs and in 25 crossbred pigs. Tissue samples were stained with Aldehyde Fuchsine. The presence of HIs was scored semi-quantitatively (HI–, HI+). We observed HIs in 48% of the purebred and in 68% of the crossbred pigs. However, only 3.3±3.1% and 3.1±4.7% of all assessed islets was hyperemic in HI+ pancreata in purebred and crossbred pigs, respectively. In both groups, significantly higher endocrine cell mass was found in the HI+ pancreata (p<0.01). When the higher endocrine cell mass was taken into account, we found significantly lower yields in the HI+ pancreata in both purebred and crossbred pigs (p=0.03 in both groups). The presence of HIs occurs frequently in porcine donor-pancreata and is associated with reduced isolation outcomes.

Key words

hyperemic - islet isolation - islet transplantation - porcine - pancreas

• References

• 1 van Suylichem PT, van Deijnen JE, Wolters GH, van Schilfgaarde R. Amount and distribution of collagen in pancreatic tissue of different species in the perspective of islet isolation procedures. Cell Transplant 1995; 4: 609-614
• 2 Ulrichs K, Bosse M, Heiser A, Eckstein V, Wacker H-H, Thiede A, Ruchholtz W. Histomorphological characteristics of the porcine pancreas as a basis for the isolation of islets of Langerhans. Xenotransplantation 1995; 2: 176-187
• 3 Socci C, Ricordi C, Davalli AM, Staudacher C, Baro P, Vertova A, Freschi M, Gavazzi F, Braghi S, Pozza G, Di Carlo V. Selection of donors significantly improves pig islet isolation yield. Horm Metab Res Suppl 1990; 25: 32-34
• 4 White SA, Hughes DP, Contractor HH, London NJ. An investigation into the distribution of different collagen types within adult and juvenile porcine pancreata. J Mol Med 1999; 77: 79-82
• 5 van Deijnen JH, Hulstaert CE, Wolters GH, van Schilfgaarde R. Significance of the peri-insular extracellular matrix for islet isolation from the pancreas of rat, dog, pig, and man. Cell Tissue Res 1992; 267: 139-146
• 6 van Deijnen JH, van Suylichem PT, Wolters GH, van Schilfgaarde R. Distribution of collagens type I, type III and type V in the pancreas of rat, dog, pig and man. Cell Tissue Res 1994; 277: 115-121
• 7 Basta G, Falorni A, Osticioli L, Brunetti P, Calafiore R. Method for mass retrieval, morphologic, and functional characterization of adult porcine islets of Langerhans: a potential nonhuman pancreatic tissue resource for xenotransplantation in insulin-dependent diabetes mellitus. J Investig Med 1995; 43: 555-566
• 8 Coleman R, Silbermann M. Erythrocytes within pancreatic B-cells of corticosteroid-treated mice. Experientia 1978; 34: 1049-1050
• 9 Imaoka M, Satoh H, Furuhama K. Age- and sex-related differences in spontaneous hemorrhage and fibrosis of the pancreatic islets in Sprague-Dawley rats. Toxicol Pathol 2007; 35: 388-394
• 10 Kaduk B, Husslein EM, Siegfried A. Morphology of the chronic toxicity of busulfan on the islets of Langerhans in the rat. Hepatogastroenterol 1987; 34: 108-112
• 11 Lucocq JM, Findlay JA. Islet organ, blood glucose and glucose tolerance of lean and obese Mongolian gerbils. A quantitative study. Cell Tissue Res 1981; 220: 623-636
• 12 Hilling DE, Töns HA, Marang-van de Mheen PJ, Baranski AG, van den Akker EK, Terpstra OT, Bouwman E. Presence of hyperemic islets in human donor-pancreata results in reduced islet isolation yield. Horm Metab Res 2011; 43: 92-99
• 13 Brissova M, Fowler MJ, Nicholson WE, Chu A, Hirshberg B, Harlan DM, Powers AC. Assessment of human pancreatic islet architecture and composition by laser scanning confocal microscopy. J Histochem Cytochem 2005; 53: 1087-1097
• 14 Cabrera O, Berman DM, Kenyon NS, Ricordi C, Berggren PO, Caicedo A. The unique cytoarchitecture of human pancreatic islets has implications for islet cell function. Proc Natl Acad Sci USA 2006; 103: 2334-2339
• 15 Moldovan S, Livingston E, Livingston E, Kleinman R, Guth P, Brunicardi FC. Glucose-induced islet hyperemia is mediated by nitric oxide. Am J Surg 1996; 171: 16-20
• 16 Grandin T. Cardiac Arrest Stunning of Livestock and Poultry. In: Fox MW, Mickley LD. eds Advances in Animal Welfare Science. The Hague: Martinus Nijhoff; 1985: 1-30
• 17 Burson DE, Hunt MC, Schafer DE, Beckwith D, Garrison JR. Effects of stunning method and time interval from stunning to exsanguination on blood splashing in pork. J Anim Sci 1983; 57: 918-921
• 18 Petersen GV, Carr DH, Davies AS, Pickett BT. The effect of different methods of electrical stunning of lambs on blood pressure and muscular activity. Meat Sci 1986; 161-165