Cryopreservation of dissociated rat pancreatic islet cells. Poster

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00023001%3A_____%2F24%3A00084524" target="_blank" >RIV/00023001:_____/24:00084524 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Cryopreservation of dissociated rat pancreatic islet cells. Poster

Popis výsledku v původním jazyce

BackgroundLong-term cultivation of isolated islets leads to dedifferentiation of endocrine cells or their death [1].Cryopreservation of whole islets is a technically demanding process [2, 3]. To preserve islet cells forlater use, we have developed a method for cryopreservation of dissociated rat pancreatic islet cells.MethodsIsolated rat pancreatic islets were dissociated into cell suspension by Accutase (20 min, RT). Somecells were used fresh and some were frozen by controlled cooling in CMRL medium supplemented with20% FBS and 10% DMSO. Frozen samples were stored in liquid nitrogen untill next usage. Fresh andthawed cells were tested for viability by Vi-CELL XR Cell Viability Analyzer (Beckman Coulter) andseeded into 96 well plate coated with extracellular matrix produced by bladder cancer cells (HTB-9),50 000 cells per well. Cultivated cells were tested for glucose stimulated insulin secretion.Subsequently, beta and alpha cell were immunolabeled to analyse their occurrence.ResultsViability of thawed cells was slightly lower than that of fresh cells, (54% versus 56%). Glucosestimulated insulin secretion was comparable in thawed and fresh cells, stimulation index was 3 and 3.5,respectively. Cultivated dissociated cells successfully adhered on the coated surface of culture plateand both beta cells and alpha cells were represented among them in similar ratio, in frozen samplesthere were 32 alpha cells per 100 beta cells and in fresh samples 37 alpha cells per 100 beta cells.ConclusionsThe results of this preliminary study show that cryopreserved dissociated rat islet cells have comparablefunctional quality to fresh cells. This approach represents a simple method for long-term preservationof islet cells.

Název v anglickém jazyce

Cryopreservation of dissociated rat pancreatic islet cells. Poster

Popis výsledku anglicky

BackgroundLong-term cultivation of isolated islets leads to dedifferentiation of endocrine cells or their death [1].Cryopreservation of whole islets is a technically demanding process [2, 3]. To preserve islet cells forlater use, we have developed a method for cryopreservation of dissociated rat pancreatic islet cells.MethodsIsolated rat pancreatic islets were dissociated into cell suspension by Accutase (20 min, RT). Somecells were used fresh and some were frozen by controlled cooling in CMRL medium supplemented with20% FBS and 10% DMSO. Frozen samples were stored in liquid nitrogen untill next usage. Fresh andthawed cells were tested for viability by Vi-CELL XR Cell Viability Analyzer (Beckman Coulter) andseeded into 96 well plate coated with extracellular matrix produced by bladder cancer cells (HTB-9),50 000 cells per well. Cultivated cells were tested for glucose stimulated insulin secretion.Subsequently, beta and alpha cell were immunolabeled to analyse their occurrence.ResultsViability of thawed cells was slightly lower than that of fresh cells, (54% versus 56%). Glucosestimulated insulin secretion was comparable in thawed and fresh cells, stimulation index was 3 and 3.5,respectively. Cultivated dissociated cells successfully adhered on the coated surface of culture plateand both beta cells and alpha cells were represented among them in similar ratio, in frozen samplesthere were 32 alpha cells per 100 beta cells and in fresh samples 37 alpha cells per 100 beta cells.ConclusionsThe results of this preliminary study show that cryopreserved dissociated rat islet cells have comparablefunctional quality to fresh cells. This approach represents a simple method for long-term preservationof islet cells.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

30202 - Endocrinology and metabolism (including diabetes, hormones)

Projekt

<a href="/cs/project/LX22NPO5104" target="_blank" >LX22NPO5104: Národní institut pro výzkum metabolických a kardiovaskulárních onemocnění</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2024

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů