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

Publisher: Public Library of Science Country of Publication: United States NLM ID: 101285081 Publication Model: eCollection Cited Medium: Internet ISSN: 1932-6203 (Electronic) Linking ISSN: 19326203 NLM ISO Abbreviation: PLoS One Subsets: MEDLINE

Original Publication: San Francisco, CA : Public Library of Science

Culture Media/*chemistry ; Imidazoles/*pharmacology ; Indoles/*pharmacology ; Islets of Langerhans/*metabolism; Animals ; Cell Survival/drug effects ; Culture Media/pharmacology ; Dose-Response Relationship, Drug ; Glucose/metabolism ; Imidazoles/metabolism ; Indoles/metabolism ; Insulin/metabolism ; Insulin Secretion/drug effects ; Islets of Langerhans Transplantation/methods ; Pancreas/metabolism ; Swine ; Tissue Culture Techniques/methods

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.
Competing Interests: The authors have declared that no competing interests exist.

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figshare 10.6084/m9.figshare.12806555

0 (Culture Media)
0 (Imidazoles)
0 (Indoles)
0 (Insulin)
0 (necrostatin-1)
IY9XDZ35W2 (Glucose)

Date Created: 20201207 Date Completed: 20210128 Latest Revision: 20210128

20240104

PMC7721208

10.1371/journal.pone.0243506

33284818