Cryopreservation of dissociated rat pancreatic islet cells. Poster
The result's identifiers
Result code in 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>
Result on the web
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DOI - Digital Object Identifier
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Alternative languages
Result language
angličtina
Original language name
Cryopreservation of dissociated rat pancreatic islet cells. Poster
Original language description
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.
Czech name
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Czech description
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Classification
Type
O - Miscellaneous
CEP classification
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OECD FORD branch
30202 - Endocrinology and metabolism (including diabetes, hormones)
Result continuities
Project
<a href="/en/project/LX22NPO5104" target="_blank" >LX22NPO5104: National Institute for Research of Metabolic and Cardiovascular Diseases</a><br>
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2024
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů