FGF-2b and h-PL Transform Duct and Non-Endocrine Human Pancreatic Cells into Endocrine Insulin Secreting Cells by Modulating Differentiating Genes
Background: Diabetes mellitus (DM) is a multifactorial disease orphan of a cure. Regenerative medicine has been proposed as novel strategy for DM therapy. Human fibroblast growth factor (FGF)-2b controls β-cell clusters via autocrine action, and human placental lactogen (hPL)-A increases functional...
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MDPI AG
2017-10-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/18/11/2234 |
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| author | Giulia Donadel Donatella Pastore David Della-Morte Barbara Capuani Marco F. Lombardo Francesca Pacifici Marco Bugliani Fabio A. Grieco Piero Marchetti Davide Lauro |
| author_facet | Giulia Donadel Donatella Pastore David Della-Morte Barbara Capuani Marco F. Lombardo Francesca Pacifici Marco Bugliani Fabio A. Grieco Piero Marchetti Davide Lauro |
| author_sort | Giulia Donadel |
| collection | DOAJ |
| description | Background: Diabetes mellitus (DM) is a multifactorial disease orphan of a cure. Regenerative medicine has been proposed as novel strategy for DM therapy. Human fibroblast growth factor (FGF)-2b controls β-cell clusters via autocrine action, and human placental lactogen (hPL)-A increases functional β-cells. We hypothesized whether FGF-2b/hPL-A treatment induces β-cell differentiation from ductal/non-endocrine precursor(s) by modulating specific genes expression. Methods: Human pancreatic ductal-cells (PANC-1) and non-endocrine pancreatic cells were treated with FGF-2b plus hPL-A at 500 ng/mL. Cytofluorimetry and Immunofluorescence have been performed to detect expression of endocrine, ductal and acinar markers. Bromodeoxyuridine incorporation and annexin-V quantified cells proliferation and apoptosis. Insulin secretion was assessed by RIA kit, and electron microscopy analyzed islet-like clusters. Results: Increase in PANC-1 duct cells de-differentiation into islet-like aggregates was observed after FGF-2b/hPL-A treatment showing ultrastructure typical of islets-aggregates. These clusters, after stimulation with FGF-2b/hPL-A, had significant (p < 0.05) increase in insulin, C-peptide, pancreatic and duodenal homeobox 1 (PDX-1), Nkx2.2, Nkx6.1, somatostatin, glucagon, and glucose transporter 2 (Glut-2), compared with control cells. Markers of PANC-1 (Cytokeratin-19, MUC-1, CA19-9) were decreased (p < 0.05). These aggregates after treatment with FGF-2b/hPL-A significantly reduced levels of apoptosis. Conclusions: FGF-2b and hPL-A are promising candidates for regenerative therapy in DM by inducing de-differentiation of stem cells modulating pivotal endocrine genes. |
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| issn | 1422-0067 |
| language | English |
| last_indexed | 2024-04-12T10:18:46Z |
| publishDate | 2017-10-01 |
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| spelling | doaj.art-0eab278443514e73b45ed8fc99c42ee42022-12-22T03:37:08ZengMDPI AGInternational Journal of Molecular Sciences1422-00672017-10-011811223410.3390/ijms18112234ijms18112234FGF-2b and h-PL Transform Duct and Non-Endocrine Human Pancreatic Cells into Endocrine Insulin Secreting Cells by Modulating Differentiating GenesGiulia Donadel0Donatella Pastore1David Della-Morte2Barbara Capuani3Marco F. Lombardo4Francesca Pacifici5Marco Bugliani6Fabio A. Grieco7Piero Marchetti8Davide Lauro9Department of Systems Medicine, University of Rome Tor Vergata, 00133 Rome, ItalyDepartment of Systems Medicine, University of Rome Tor Vergata, 00133 Rome, ItalyDepartment of Systems Medicine, University of Rome Tor Vergata, 00133 Rome, ItalyDepartment of Systems Medicine, University of Rome Tor Vergata, 00133 Rome, ItalyAgenzia regionale per la protezione ambientale (ARPA) Lazio, Sezione di Roma, 00173 Rome, ItalyDepartment of Systems Medicine, University of Rome Tor Vergata, 00133 Rome, ItalyEndocrinology and Metabolism of Transplantation, Azienda Ospedaliero-Universitaria (A.O.U.) Pisana, 56126 Pisa, ItalyDepartment of Medicine, Surgery and Neuroscience, University of Siena, 53100 Siena, ItalyEndocrinology and Metabolism of Transplantation, Azienda Ospedaliero-Universitaria (A.O.U.) Pisana, 56126 Pisa, ItalyDepartment of Systems Medicine, University of Rome Tor Vergata, 00133 Rome, ItalyBackground: Diabetes mellitus (DM) is a multifactorial disease orphan of a cure. Regenerative medicine has been proposed as novel strategy for DM therapy. Human fibroblast growth factor (FGF)-2b controls β-cell clusters via autocrine action, and human placental lactogen (hPL)-A increases functional β-cells. We hypothesized whether FGF-2b/hPL-A treatment induces β-cell differentiation from ductal/non-endocrine precursor(s) by modulating specific genes expression. Methods: Human pancreatic ductal-cells (PANC-1) and non-endocrine pancreatic cells were treated with FGF-2b plus hPL-A at 500 ng/mL. Cytofluorimetry and Immunofluorescence have been performed to detect expression of endocrine, ductal and acinar markers. Bromodeoxyuridine incorporation and annexin-V quantified cells proliferation and apoptosis. Insulin secretion was assessed by RIA kit, and electron microscopy analyzed islet-like clusters. Results: Increase in PANC-1 duct cells de-differentiation into islet-like aggregates was observed after FGF-2b/hPL-A treatment showing ultrastructure typical of islets-aggregates. These clusters, after stimulation with FGF-2b/hPL-A, had significant (p < 0.05) increase in insulin, C-peptide, pancreatic and duodenal homeobox 1 (PDX-1), Nkx2.2, Nkx6.1, somatostatin, glucagon, and glucose transporter 2 (Glut-2), compared with control cells. Markers of PANC-1 (Cytokeratin-19, MUC-1, CA19-9) were decreased (p < 0.05). These aggregates after treatment with FGF-2b/hPL-A significantly reduced levels of apoptosis. Conclusions: FGF-2b and hPL-A are promising candidates for regenerative therapy in DM by inducing de-differentiation of stem cells modulating pivotal endocrine genes.https://www.mdpi.com/1422-0067/18/11/2234pancreatic β cellscellular differentiationinsulin releaseregenerative medicinediabetes mellitus |
| spellingShingle | Giulia Donadel Donatella Pastore David Della-Morte Barbara Capuani Marco F. Lombardo Francesca Pacifici Marco Bugliani Fabio A. Grieco Piero Marchetti Davide Lauro FGF-2b and h-PL Transform Duct and Non-Endocrine Human Pancreatic Cells into Endocrine Insulin Secreting Cells by Modulating Differentiating Genes International Journal of Molecular Sciences pancreatic β cells cellular differentiation insulin release regenerative medicine diabetes mellitus |
| title | FGF-2b and h-PL Transform Duct and Non-Endocrine Human Pancreatic Cells into Endocrine Insulin Secreting Cells by Modulating Differentiating Genes |
| title_full | FGF-2b and h-PL Transform Duct and Non-Endocrine Human Pancreatic Cells into Endocrine Insulin Secreting Cells by Modulating Differentiating Genes |
| title_fullStr | FGF-2b and h-PL Transform Duct and Non-Endocrine Human Pancreatic Cells into Endocrine Insulin Secreting Cells by Modulating Differentiating Genes |
| title_full_unstemmed | FGF-2b and h-PL Transform Duct and Non-Endocrine Human Pancreatic Cells into Endocrine Insulin Secreting Cells by Modulating Differentiating Genes |
| title_short | FGF-2b and h-PL Transform Duct and Non-Endocrine Human Pancreatic Cells into Endocrine Insulin Secreting Cells by Modulating Differentiating Genes |
| title_sort | fgf 2b and h pl transform duct and non endocrine human pancreatic cells into endocrine insulin secreting cells by modulating differentiating genes |
| topic | pancreatic β cells cellular differentiation insulin release regenerative medicine diabetes mellitus |
| url | https://www.mdpi.com/1422-0067/18/11/2234 |
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