The role of CMV in the pathology and etiology of islet graft failure

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Isolation of primary beta-cells from other cells within in the pancreatic islets is of importance for many fields of islet research. However, up to now, no satisfactory method has been developed that gained high numbers of viable beta-cells, without considerable alpha-cell contamination. In this study we investigated whether rat beta-cells can be isolated from non-beta endocrine cells by manipulating the Flavin Adenine Dinucleotide (FAD) and Nicotinamide Adenine Dinucleotide phosphate (NAD(P)H) autofluorescence.
Beta-cells were isolated from dispersed islets by flow cytometry, based on their high FAD and NAD(P)H flurorescence. To improve beta-cell yield and purity, the cellular FAD and NAD(P)H content was altered by pre-incubation in culture media containing varying amounts of D-glucose and amino acids.
Manipulation of the cellular FAD and NAD(P)H fluorescence improves beta-cell yield and purity after sorting. This method is also a fast and reliable method to measure beta-cell functional viability. A conceivable application is assessing beta-cell viability before transplantation.

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