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dc.contributor.authorLemos, Natália Emerimpt_BR
dc.contributor.authorDieter, Cristinept_BR
dc.contributor.authorFarias, Mariela Graneropt_BR
dc.contributor.authorRheinheimer, Jakelinept_BR
dc.contributor.authorSouza, Bianca Marmontel dept_BR
dc.contributor.authorBauer, Andrea Carlapt_BR
dc.contributor.authorCrispim, Daisypt_BR
dc.date.accessioned2022-01-07T04:27:00Zpt_BR
dc.date.issued2021pt_BR
dc.identifier.issn2357-9730pt_BR
dc.identifier.urihttp://hdl.handle.net/10183/233833pt_BR
dc.description.abstractIntroduction: The success of islet transplantation for patients with unstable type 1 diabetes mellitus depends, in part, on the number of isolated islets and their quality, which is assessed by functional and viability tests. The test currently employed to evaluate islet viability, used by the Collaborative Islet Transplant Registry to release products for transplantation, is fluorescein diacetate/propidium iodide (FDA/PI) staining. However, the efficacy of this method relies on researcher experience; in this context, a quantitative method may be useful. The aim of this study was to compare islet viability as assessed by flow cytometry and the FDA/PI assay. Methods: Viability was analyzed in islets isolated from 10 male Wistar rats. Upon FDA/PI staining, 50 islets from each animal were analyzed under fluorescence microscopy by two well-trained researchers. For flow cytometry, islets were dispersed and 100 000 single cells were incubated with the 7-amino-actinomycin D (7AAD) fluorophore (dyes necrotic and late apoptotic cells) and the Annexin V-APC antibody (marks early apoptotic cells). Results: A moderate correlation was found between techniques (r = 0.6; p = 0.047). The mean islet viability measured by flow cytometry was higher than that estimated using FDA/PI staining (95.5 ± 1.4% vs 89.5 ± 5.0%; p = 0.002). Conclusions: Although flow cytometry is more expensive and time-consuming than FDA/PI staining, it is a quantitative technique with greater reproducibility that is less subject to inter-observer variability than FDA/PI. Therefore, flow cytometry appears to be the technique of choice when aiming for a more precise determination of islet viability.en
dc.format.mimetypeapplication/pdfpt_BR
dc.language.isoengpt_BR
dc.relation.ispartofClinical and biomedical research. Porto Alegre. Vol. 41, no. 4 (2021), p. 325-331pt_BR
dc.rightsOpen Accessen
dc.subjectIslet transplantationen
dc.subjectTransplante das ilhotas pancreáticaspt_BR
dc.subjectIslet isolationen
dc.subjectIlhotas pancreáticaspt_BR
dc.subjectViabilityen
dc.subjectSeparação celularpt_BR
dc.subjectSobrevivência celularpt_BR
dc.subjectFlow cytometryen
dc.subjectCitometria de fluxopt_BR
dc.titleComparison of two techniques for assessing pancreatic islet viability : flow cytometry and fluorescein diacetate/propidium iodide stainingpt_BR
dc.typeArtigo de periódicopt_BR
dc.identifier.nrb001135178pt_BR
dc.type.originNacionalpt_BR


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