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Pluripotency-associated stem cell marker expression in proliferative cell cultures derived from adult human pancreas
Lookup NU author(s)
Dr Michael White
Dr Graham Holliman
Dr Ali Aldibbiat
Professor James Shaw
White MG, Al-Turaifi HR, Holliman GN, Aldibbiat A, Mahmoud A, Shaw JAM
Journal of Endocrinology
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The source of new beta-cells in adult human pancreas remains incompletely elucidated with recent studies in rodents providing evidence for neogenesis from progenitor cells in addition to self-replication. The aim of this study was to investigate expression of pluripotency-associated stem cell markers in proliferative cultures derived from adult human pancreas.Human pancreatic tissue was obtained from deceased donors following ethical approval and relative consent. Islet-enriched fraction was separated from the retrieved organ by digestion and density gradient centrifugation. Dissociated cells were seeded in adherent culture forming proliferative 'islet survivor cells'(ISCs). These were characterized at fifth passage by RT-PCR, immunofluorescence staining, FACS, Western blot and transfection studies with an OCT4 promoter-driven reporter.Nuclear expression of the pluripotency-associated stem cell marker complex OCT4 / SOX2 / NANOG was confirmed in ISCs. The phenotype constituted approximately 8% of the overall population. OCT4 biosynthesis was confirmed by Western blot and activation of an exogenous OCT4 promoter.Co-expression of pluripotency-associated markers has been confirmed in proliferative primary cells derived from adult human pancreas. Further studies are required to elucidate whether these cells possess functional stem cell characteristics and to assess potential for differentiation into pancreatic cell lineages including new beta-cells.
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