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Transfection of Peripheral Blood Monocytes with SOX2 Enhances Multipotency, Proliferation, and Redifferentiation into Neohepatocytes and Insulin-Producing Cells

Following a several-day incubation in medium containing IL-3 and M-CSF to generate a more plastic intermediate “reprogrammed multipotent cells of monocytic origin (RMCMO),” peripheral blood mononuclear cells (PBMCs) can be efficiently converted to hepatocyte-like cells (neohepatocytes) and insulin-p...

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Autores principales: Hyder, Ayman, Ehnert, Sabrina, Fändrich, Fred, Ungefroren, Hendrik
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6193350/
https://www.ncbi.nlm.nih.gov/pubmed/30402109
http://dx.doi.org/10.1155/2018/4271875
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author Hyder, Ayman
Ehnert, Sabrina
Fändrich, Fred
Ungefroren, Hendrik
author_facet Hyder, Ayman
Ehnert, Sabrina
Fändrich, Fred
Ungefroren, Hendrik
author_sort Hyder, Ayman
collection PubMed
description Following a several-day incubation in medium containing IL-3 and M-CSF to generate a more plastic intermediate “reprogrammed multipotent cells of monocytic origin (RMCMO),” peripheral blood mononuclear cells (PBMCs) can be efficiently converted to hepatocyte-like cells (neohepatocytes) and insulin-producing cells. However, continuous efforts are devoted to enhancing the proliferative capacity of these multipotent cells while maintaining or further increasing their redifferentiation potential. In the present work, PBMCs were transfected with one pluripotency gene (SOX2) and the resulting RMCMO compared to standard RMCMO with respect to cell viability, proliferative activity, and redifferentiation potential. Ectopic SOX2 expression increased the number of viable RMCMO, activated cell cycle genes, and enhanced proliferation as shown by quantitative RT-PCR and Ki67 immunofluorescent staining, respectively. Redifferentiation of RMCMO derived from SOX2-transfected PBMCs to neohepatocytes was more complete in comparison to control cells as revealed by higher urea and glucose secretion, increased activity of cytochrome P450 isoforms, and a phase II enzyme, while the same was true for insulin-producing cells as assessed by the expression of INS, PDX1, and GLUT2 and glucose-stimulated insulin secretion. Our results indicate that SOX2 transfection increases both multipotency and proliferation of RMCMO, eventually allowing production of neohepatocytes and insulin-producing cells of higher quality and quantity for transplantation purposes.
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spelling pubmed-61933502018-11-06 Transfection of Peripheral Blood Monocytes with SOX2 Enhances Multipotency, Proliferation, and Redifferentiation into Neohepatocytes and Insulin-Producing Cells Hyder, Ayman Ehnert, Sabrina Fändrich, Fred Ungefroren, Hendrik Stem Cells Int Research Article Following a several-day incubation in medium containing IL-3 and M-CSF to generate a more plastic intermediate “reprogrammed multipotent cells of monocytic origin (RMCMO),” peripheral blood mononuclear cells (PBMCs) can be efficiently converted to hepatocyte-like cells (neohepatocytes) and insulin-producing cells. However, continuous efforts are devoted to enhancing the proliferative capacity of these multipotent cells while maintaining or further increasing their redifferentiation potential. In the present work, PBMCs were transfected with one pluripotency gene (SOX2) and the resulting RMCMO compared to standard RMCMO with respect to cell viability, proliferative activity, and redifferentiation potential. Ectopic SOX2 expression increased the number of viable RMCMO, activated cell cycle genes, and enhanced proliferation as shown by quantitative RT-PCR and Ki67 immunofluorescent staining, respectively. Redifferentiation of RMCMO derived from SOX2-transfected PBMCs to neohepatocytes was more complete in comparison to control cells as revealed by higher urea and glucose secretion, increased activity of cytochrome P450 isoforms, and a phase II enzyme, while the same was true for insulin-producing cells as assessed by the expression of INS, PDX1, and GLUT2 and glucose-stimulated insulin secretion. Our results indicate that SOX2 transfection increases both multipotency and proliferation of RMCMO, eventually allowing production of neohepatocytes and insulin-producing cells of higher quality and quantity for transplantation purposes. Hindawi 2018-10-04 /pmc/articles/PMC6193350/ /pubmed/30402109 http://dx.doi.org/10.1155/2018/4271875 Text en Copyright © 2018 Ayman Hyder et al. http://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
Hyder, Ayman
Ehnert, Sabrina
Fändrich, Fred
Ungefroren, Hendrik
Transfection of Peripheral Blood Monocytes with SOX2 Enhances Multipotency, Proliferation, and Redifferentiation into Neohepatocytes and Insulin-Producing Cells
title Transfection of Peripheral Blood Monocytes with SOX2 Enhances Multipotency, Proliferation, and Redifferentiation into Neohepatocytes and Insulin-Producing Cells
title_full Transfection of Peripheral Blood Monocytes with SOX2 Enhances Multipotency, Proliferation, and Redifferentiation into Neohepatocytes and Insulin-Producing Cells
title_fullStr Transfection of Peripheral Blood Monocytes with SOX2 Enhances Multipotency, Proliferation, and Redifferentiation into Neohepatocytes and Insulin-Producing Cells
title_full_unstemmed Transfection of Peripheral Blood Monocytes with SOX2 Enhances Multipotency, Proliferation, and Redifferentiation into Neohepatocytes and Insulin-Producing Cells
title_short Transfection of Peripheral Blood Monocytes with SOX2 Enhances Multipotency, Proliferation, and Redifferentiation into Neohepatocytes and Insulin-Producing Cells
title_sort transfection of peripheral blood monocytes with sox2 enhances multipotency, proliferation, and redifferentiation into neohepatocytes and insulin-producing cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6193350/
https://www.ncbi.nlm.nih.gov/pubmed/30402109
http://dx.doi.org/10.1155/2018/4271875
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