Cargando…

Dose-dependent effects of necrostatin-1 supplementation to tissue culture media of young porcine islets

Previous studies have shown that necrostatin-1 (Nec-1) supplementation improved the viability of murine islets following exposure to nitric oxide, increased the survival of human islets during hypoxic culture, and augmented the maturation of pre-weaned porcine islets (PPIs) after 7 days of tissue cu...

Descripción completa

Detalles Bibliográficos
Autores principales: Lau, Hien, Corrales, Nicole, Rodriguez, Samuel, Luong, Colleen, Mohammadi, Mohammadreza, Khosrawipour, Veria, Li, Shiri, Alexander, Michael, de Vos, Paul, Lakey, Jonathan R. T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7721208/
https://www.ncbi.nlm.nih.gov/pubmed/33284818
http://dx.doi.org/10.1371/journal.pone.0243506
_version_ 1783619996547547136
author Lau, Hien
Corrales, Nicole
Rodriguez, Samuel
Luong, Colleen
Mohammadi, Mohammadreza
Khosrawipour, Veria
Li, Shiri
Alexander, Michael
de Vos, Paul
Lakey, Jonathan R. T.
author_facet Lau, Hien
Corrales, Nicole
Rodriguez, Samuel
Luong, Colleen
Mohammadi, Mohammadreza
Khosrawipour, Veria
Li, Shiri
Alexander, Michael
de Vos, Paul
Lakey, Jonathan R. T.
author_sort Lau, Hien
collection PubMed
description Previous studies have shown that necrostatin-1 (Nec-1) supplementation improved the viability of murine islets following exposure to nitric oxide, increased the survival of human islets during hypoxic culture, and augmented the maturation of pre-weaned porcine islets (PPIs) after 7 days of tissue culture. A limitation of these studies is that only one concentration of Nec-1 was used, and no studies have determined the optimal dose of Nec-1 for PPIs. Thus, the present study examined the effects of Nec-1 on PPIs at four different doses—0, 25, 50, 100, and 200 μM—after 7 days of tissue culture when supplemented on day 3. PPIs were isolated from pancreata of pre-weaned Yorkshire piglets (8–15 days old) and cultured in a specific islet maturation media added with Nec-1 on day 3 of tissue culture at 4 different doses—0, 25, 50, 100, and 200 μM (n = 6 for each dose). After 7 days of tissue culture, islets were assessed for recovery, viability, endocrine cellular content, GLUT2 expression in beta cells, and insulin secretion after glucose challenge. Nec-1 did not affect the viability of both intact islets and dissociated islets cells during tissue culture regardless of doses. Islets cultured in media supplemented with Nec-1 at 100 μM, but not 25, 50, or 200 μM, had a significantly higher recovery, composition of endocrine cells, GLUT2 expression in beta cells, and insulin secretion capacity than control islets cultured in media without Nec-1 supplementation. Moreover, culturing islets in 200 μM Nec-1 supplemented media not only failed to improve the insulin release but resulted in a lower glucose-induced insulin stimulation index compared to islets cultured in media added with 100 μM Nec-1. Xenotransplantation using porcine islets continues to demonstrate scientific advances to justify this area of research. Our findings indicate that Nec-1 supplementation at 100 μM was most effective to enhance the in vitro maturation of PPIs during tissue culture.
format Online
Article
Text
id pubmed-7721208
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-77212082020-12-15 Dose-dependent effects of necrostatin-1 supplementation to tissue culture media of young porcine islets Lau, Hien Corrales, Nicole Rodriguez, Samuel Luong, Colleen Mohammadi, Mohammadreza Khosrawipour, Veria Li, Shiri Alexander, Michael de Vos, Paul Lakey, Jonathan R. T. PLoS One Research Article Previous studies have shown that necrostatin-1 (Nec-1) supplementation improved the viability of murine islets following exposure to nitric oxide, increased the survival of human islets during hypoxic culture, and augmented the maturation of pre-weaned porcine islets (PPIs) after 7 days of tissue culture. A limitation of these studies is that only one concentration of Nec-1 was used, and no studies have determined the optimal dose of Nec-1 for PPIs. Thus, the present study examined the effects of Nec-1 on PPIs at four different doses—0, 25, 50, 100, and 200 μM—after 7 days of tissue culture when supplemented on day 3. PPIs were isolated from pancreata of pre-weaned Yorkshire piglets (8–15 days old) and cultured in a specific islet maturation media added with Nec-1 on day 3 of tissue culture at 4 different doses—0, 25, 50, 100, and 200 μM (n = 6 for each dose). After 7 days of tissue culture, islets were assessed for recovery, viability, endocrine cellular content, GLUT2 expression in beta cells, and insulin secretion after glucose challenge. Nec-1 did not affect the viability of both intact islets and dissociated islets cells during tissue culture regardless of doses. Islets cultured in media supplemented with Nec-1 at 100 μM, but not 25, 50, or 200 μM, had a significantly higher recovery, composition of endocrine cells, GLUT2 expression in beta cells, and insulin secretion capacity than control islets cultured in media without Nec-1 supplementation. Moreover, culturing islets in 200 μM Nec-1 supplemented media not only failed to improve the insulin release but resulted in a lower glucose-induced insulin stimulation index compared to islets cultured in media added with 100 μM Nec-1. Xenotransplantation using porcine islets continues to demonstrate scientific advances to justify this area of research. Our findings indicate that Nec-1 supplementation at 100 μM was most effective to enhance the in vitro maturation of PPIs during tissue culture. Public Library of Science 2020-12-07 /pmc/articles/PMC7721208/ /pubmed/33284818 http://dx.doi.org/10.1371/journal.pone.0243506 Text en © 2020 Lau et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Lau, Hien
Corrales, Nicole
Rodriguez, Samuel
Luong, Colleen
Mohammadi, Mohammadreza
Khosrawipour, Veria
Li, Shiri
Alexander, Michael
de Vos, Paul
Lakey, Jonathan R. T.
Dose-dependent effects of necrostatin-1 supplementation to tissue culture media of young porcine islets
title Dose-dependent effects of necrostatin-1 supplementation to tissue culture media of young porcine islets
title_full Dose-dependent effects of necrostatin-1 supplementation to tissue culture media of young porcine islets
title_fullStr Dose-dependent effects of necrostatin-1 supplementation to tissue culture media of young porcine islets
title_full_unstemmed Dose-dependent effects of necrostatin-1 supplementation to tissue culture media of young porcine islets
title_short Dose-dependent effects of necrostatin-1 supplementation to tissue culture media of young porcine islets
title_sort dose-dependent effects of necrostatin-1 supplementation to tissue culture media of young porcine islets
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7721208/
https://www.ncbi.nlm.nih.gov/pubmed/33284818
http://dx.doi.org/10.1371/journal.pone.0243506
work_keys_str_mv AT lauhien dosedependenteffectsofnecrostatin1supplementationtotissueculturemediaofyoungporcineislets
AT corralesnicole dosedependenteffectsofnecrostatin1supplementationtotissueculturemediaofyoungporcineislets
AT rodriguezsamuel dosedependenteffectsofnecrostatin1supplementationtotissueculturemediaofyoungporcineislets
AT luongcolleen dosedependenteffectsofnecrostatin1supplementationtotissueculturemediaofyoungporcineislets
AT mohammadimohammadreza dosedependenteffectsofnecrostatin1supplementationtotissueculturemediaofyoungporcineislets
AT khosrawipourveria dosedependenteffectsofnecrostatin1supplementationtotissueculturemediaofyoungporcineislets
AT lishiri dosedependenteffectsofnecrostatin1supplementationtotissueculturemediaofyoungporcineislets
AT alexandermichael dosedependenteffectsofnecrostatin1supplementationtotissueculturemediaofyoungporcineislets
AT devospaul dosedependenteffectsofnecrostatin1supplementationtotissueculturemediaofyoungporcineislets
AT lakeyjonathanrt dosedependenteffectsofnecrostatin1supplementationtotissueculturemediaofyoungporcineislets