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In Vitro Proliferation of Porcine Pancreatic Islet Cells for β-Cell Therapy Applications

β-Cell replacement through transplantation is the only curative treatment to establish a long-term stable euglycemia in diabetic patients. Owing to the shortage of donor tissue, attempts are being made to develop alternative sources of insulin-secreting cells. Stem cells differentiation and reprogra...

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Autores principales: Niu, Guoguang, McQuilling, John P., Zhou, Yu, Opara, Emmanuel C., Orlando, Giuseppe, Soker, Shay
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5168554/
https://www.ncbi.nlm.nih.gov/pubmed/28050568
http://dx.doi.org/10.1155/2016/5807876
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author Niu, Guoguang
McQuilling, John P.
Zhou, Yu
Opara, Emmanuel C.
Orlando, Giuseppe
Soker, Shay
author_facet Niu, Guoguang
McQuilling, John P.
Zhou, Yu
Opara, Emmanuel C.
Orlando, Giuseppe
Soker, Shay
author_sort Niu, Guoguang
collection PubMed
description β-Cell replacement through transplantation is the only curative treatment to establish a long-term stable euglycemia in diabetic patients. Owing to the shortage of donor tissue, attempts are being made to develop alternative sources of insulin-secreting cells. Stem cells differentiation and reprograming as well as isolating pancreatic progenitors from different sources are some examples; however, no approach has yet yielded a clinically relevant solution. Dissociated islet cells that are cultured in cell numbers by in vitro proliferation provide a promising platform for redifferentiation towards β-cells phenotype. In this study, we cultured islet-derived cells in vitro and examined the expression of β-cell genes during the proliferation. Islets were isolated from porcine pancreases and enzymatically digested to dissociate the component cells. The cells proliferated well in tissue culture plates and were subcultured for no more than 5 passages. Only 10% of insulin expression, as measured by PCR, was preserved in each passage. High glucose media enhanced insulin expression by about 4–18 fold, suggesting a glucose-dependent effect in the proliferated islet-derived cells. The islet-derived cells also expressed other pancreatic genes such as Pdx1, NeuroD, glucagon, and somatostatin. Taken together, these results indicate that pancreatic islet-derived cells, proliferated in vitro, retained the expression capacity for key pancreatic genes, thus suggesting that the cells may be redifferentiated into insulin-secreting β-like cells.
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spelling pubmed-51685542017-01-03 In Vitro Proliferation of Porcine Pancreatic Islet Cells for β-Cell Therapy Applications Niu, Guoguang McQuilling, John P. Zhou, Yu Opara, Emmanuel C. Orlando, Giuseppe Soker, Shay J Diabetes Res Research Article β-Cell replacement through transplantation is the only curative treatment to establish a long-term stable euglycemia in diabetic patients. Owing to the shortage of donor tissue, attempts are being made to develop alternative sources of insulin-secreting cells. Stem cells differentiation and reprograming as well as isolating pancreatic progenitors from different sources are some examples; however, no approach has yet yielded a clinically relevant solution. Dissociated islet cells that are cultured in cell numbers by in vitro proliferation provide a promising platform for redifferentiation towards β-cells phenotype. In this study, we cultured islet-derived cells in vitro and examined the expression of β-cell genes during the proliferation. Islets were isolated from porcine pancreases and enzymatically digested to dissociate the component cells. The cells proliferated well in tissue culture plates and were subcultured for no more than 5 passages. Only 10% of insulin expression, as measured by PCR, was preserved in each passage. High glucose media enhanced insulin expression by about 4–18 fold, suggesting a glucose-dependent effect in the proliferated islet-derived cells. The islet-derived cells also expressed other pancreatic genes such as Pdx1, NeuroD, glucagon, and somatostatin. Taken together, these results indicate that pancreatic islet-derived cells, proliferated in vitro, retained the expression capacity for key pancreatic genes, thus suggesting that the cells may be redifferentiated into insulin-secreting β-like cells. Hindawi Publishing Corporation 2016 2016-12-06 /pmc/articles/PMC5168554/ /pubmed/28050568 http://dx.doi.org/10.1155/2016/5807876 Text en Copyright © 2016 Guoguang Niu et al. https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Niu, Guoguang
McQuilling, John P.
Zhou, Yu
Opara, Emmanuel C.
Orlando, Giuseppe
Soker, Shay
In Vitro Proliferation of Porcine Pancreatic Islet Cells for β-Cell Therapy Applications
title In Vitro Proliferation of Porcine Pancreatic Islet Cells for β-Cell Therapy Applications
title_full In Vitro Proliferation of Porcine Pancreatic Islet Cells for β-Cell Therapy Applications
title_fullStr In Vitro Proliferation of Porcine Pancreatic Islet Cells for β-Cell Therapy Applications
title_full_unstemmed In Vitro Proliferation of Porcine Pancreatic Islet Cells for β-Cell Therapy Applications
title_short In Vitro Proliferation of Porcine Pancreatic Islet Cells for β-Cell Therapy Applications
title_sort in vitro proliferation of porcine pancreatic islet cells for β-cell therapy applications
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5168554/
https://www.ncbi.nlm.nih.gov/pubmed/28050568
http://dx.doi.org/10.1155/2016/5807876
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