Polski merkuriusz lekarski : organ Polskiego Towarzystwa Lekarskiego · 2008
[Islet isolation outcome is influenced by pancreas preparation method].
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Pokrywczyńska Marta, Drewa Tomasz, Cieślak Zaneta
Аннотация
UNLABELLED: Pancreatic islet transplantation is a treatment method for type I diabetes. Its outcome is influenced by numerous factors, islet quantity and function being important ones of them.
THE AIM OF THIS STUDY: was to estimate the influence of pancreas preparation method on the outcome of islet isolation in rat.
MATERIAL AND METHODS: 6 pancreata harvested from Lewis rats were used in this research. Pancreatic duct was cannulated and pancreas was injected with 1 mg/ml collagenase P solution (Sigma) and then excised. After cutting into smaller fragments, it was digested in collagenase P solution for 15-20 min. Enzyme activity was then stopped by adding dilution medium. Heterogenous cell suspension was centrifuged in density gradient (Gradisol) to isolate islets. Pancreatic islets were collected and islet equivalent was calculated. Islet purity degree was estimated as islet cells to all cells, including exocrine, ratio. Islet viability was estimated using propidium iodide and fluorescein diacetate staining. Photographic documentation was made.
RESULTS: Proper islet morphology, highest number and viability was obtained when pancreas was excised properly (isolation 3 and 4).
CONCLUSION: Pancreas preparation method is one of which influences on islet isolation outcome.
THE AIM OF THIS STUDY: was to estimate the influence of pancreas preparation method on the outcome of islet isolation in rat.
MATERIAL AND METHODS: 6 pancreata harvested from Lewis rats were used in this research. Pancreatic duct was cannulated and pancreas was injected with 1 mg/ml collagenase P solution (Sigma) and then excised. After cutting into smaller fragments, it was digested in collagenase P solution for 15-20 min. Enzyme activity was then stopped by adding dilution medium. Heterogenous cell suspension was centrifuged in density gradient (Gradisol) to isolate islets. Pancreatic islets were collected and islet equivalent was calculated. Islet purity degree was estimated as islet cells to all cells, including exocrine, ratio. Islet viability was estimated using propidium iodide and fluorescein diacetate staining. Photographic documentation was made.
RESULTS: Proper islet morphology, highest number and viability was obtained when pancreas was excised properly (isolation 3 and 4).
CONCLUSION: Pancreas preparation method is one of which influences on islet isolation outcome.