In Vitro and in Vivo Viability Assessment of Unpurified Pancreatic Islet Tissue

Rotational culture and integration with amniotic stem cells reduce porcine islet immunoreactivity in vitro and slow xeno‐rejection in a murine model of islet transplantation

“…Isolated islets preparations were found to be viable with Trypan Blue scores of greater than 75% (mean score = 22.533 ± 0.128/30) . Although prone to fragmentation during the isolation process (Figure A), porcine islets re‐aggregated during RCCS culture, as they formed constructs of 150‐200 µm in diameter over a period of 48 hours (Figure B).…”

“…Purified islets were stained with dithizone (500 µg/mL), counted and converted to islet equivalents (IEQ; 1IEQ equivalent to an islet measuring 150 µm in diameter). Islet cell viability was confirmed by Trypan Blue (0.4% v/v) exclusion . Islets were re‐suspended in Medium 199 (M199) supplemented with 10% porcine serum, 1% non‐essential amino acids, 3 mmol/L L‐glutamine and 10 µmol/L nicotinamide (vitamin B3) with anti‐microbial agents; 100 U/mL penicillin, 100 µg/mL streptomycin, 10 µg/mL amphotericin B until use in the 3‐D culture studies.…”

A Metabolomic Approach (1H HRMAS NMR Spectroscopy) Supported by Histology to Study Early Post-transplantation Responses in Islet-transplanted Livers

“…It appeared that both lactate and the lactate/alanine ratio are good markers of islet quality and oxygenation status, as they increase during hypoxia 15 , and that phosphocholine and mobile lipids are markers of apoptosis 15 . According to Hesse et al ., the phosphate diester/phosphate monoester ratio is decreased in damaged and necrotic islets, and therefore is an indicator of islet viability prior to transplantation 16 . In order to detect islet graft rejection, metabolomics was used to evaluate the role of nitric oxide (NO) metabolites as markers of the early phase of islet rejection after transplantation 17 .…”