Pluripotent stem cells isolated from human amniotic fluid and differentiation into pancreatic beta-cells.
Human amniotic fluid (HAF) contains multipotent stem cells [amniotic fluid-derived stem cells (AFSC)] which can differentiate into a variety of different cell types. Recently, we demonstrated that obestatin, a peptide encoded by the ghrelin gene, exerts anti-apoptotic effects in pancreatic beta-cells and human islets and increases the expression of genes involved in beta-cells differentiation. We investigated whether: 1) AFSC would differentiate into pancreatic beta-cells and 2) obestatin would increase beta-cells differentiation from AFSC. Fluorescence-activated cell sorting analysis and immunocytochemical staining showed the presence of mesenchymal and endothelial markers in AFSC. Real-time PCR evidenced the expression of Octamer binding transcription factor 4 (OCT-4), a marker of pluripotency, during the early differentiation phase. However, the beta-cells differentiation marker duodenal homeobox factor-1 (PDX-1) could not be detected. Obestatin increased OCT-4 expression but had no effect on beta-cells differentiation. These results suggest that, at least under the experimental conditions used in this study, AFSC do not differentiate into beta-cells and obestatin has no additional effect.
Trovato
L
L
Division of Endocrinology and Metabolism, University of Turin, 10126 Turin, Italy.
De Fazio
R
R
Annunziata
M
M
Sdei
S
S
Favaro
E
E
Ponti
R
R
Marozio
L
L
Ghigo
E
E
Benedetto
C
C
Granata
R
R
D000328:Adult; D000653:Amniotic Fluid / Q000166:cytology*; D002454:Cell Differentiation / Q000187:drug effects; D002469:Cell Separation; D002478:Cells, Cultured; D005260:Female; D018507:Gene Expression Regulation, Developmental; D054439:Ghrelin / Q000494:pharmacology; D018398:Homeodomain Proteins / Q000096:biosynthesis; D006801:Humans; D050417:Insulin-Secreting Cells / Q000166:cytology*; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects; D011247:Pregnancy; D015534:Trans-Activators / Q000096:biosynthesis
D054439:Ghrelin; D018398:Homeodomain Proteins; D050814:Octamer Transcription Factor-3; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein