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Generation and Characterization of Induced Pluripotent Stem Cells from Mononuclear Cells in Schizophrenic Patients
OBJECTIVE: Schizophrenia (SZ) is a mental disorder in which psychotic symptoms are the main problem. The pathogenesis of SZ is not fully understood, partly because of limitations in current disease models and technology. The development of induced pluripotent stem cell (iPSC) technology has opened u...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Royan Institute
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6397609/ https://www.ncbi.nlm.nih.gov/pubmed/30825289 http://dx.doi.org/10.22074/cellj.2019.5871 |
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author | Liu, Qing Du, Jiang Fan, Jinyu Li, Wenqiang Guo, Weiyun Feng, Huigen Lin, Juntang |
author_facet | Liu, Qing Du, Jiang Fan, Jinyu Li, Wenqiang Guo, Weiyun Feng, Huigen Lin, Juntang |
author_sort | Liu, Qing |
collection | PubMed |
description | OBJECTIVE: Schizophrenia (SZ) is a mental disorder in which psychotic symptoms are the main problem. The pathogenesis of SZ is not fully understood, partly because of limitations in current disease models and technology. The development of induced pluripotent stem cell (iPSC) technology has opened up the possibility of elucidating disease mechanisms in neurodegenerative diseases. Here, we aimed to obtain iPSCs from peripheral blood mononuclear cells (PBMCs) of normal and schizophrenic individuals and analyze the inflammatory response in these iPSCs. MATERIALS AND METHODS: In this experimental study, we isolated PBMCs from whole blood of healthy individuals and SZ patients and reprogrammed them into iPSCs by transfection of recombinant lentiviruses that contained Yamanaka factors (Oct4, Sox2, Klf4 and c-Myc). We calculated the numbers of iPSC clones and stained them with alkaline phosphatase (ALP), Nanog, SSEA4, Nestin, Vimentin, and AFP to confirm their efficiency and pluripotency. The iPSCs were analyzed by real-time quantitative polymerase chain reaction (qRT-PCR) for the expressions of inflammatory factors. RESULTS: iPSCs from schizophrenic patients (SZ-iPSCs) exhibited typical morphology and highly expressed pluripotent markers. These iPSCs retained their normal karyotype and differentiated in vitro to form embryoid bodies (EBs) that expressed markers of all 3 germ layers. However, iPSCs from the SZ-iPSCs group had a weak capacity to differentiate into ectoderm compared to the normal iPSCs (Con-iPSC). An elevated, stronger inflammatory response existed in iPSCs from schizophrenic individuals. CONCLUSION: We successfully obtained iPSCs from PBMCs of schizophrenic patients without genetic operation and analyzed the expressions of pluripotent markers and inflammatory factors between the Con-iPSC and SZ-iPSC groups. Taken together, our results may assist to explain the pathogenesis of SZ and develop new strategies for clinical diagnosis and treatment. |
format | Online Article Text |
id | pubmed-6397609 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Royan Institute |
record_format | MEDLINE/PubMed |
spelling | pubmed-63976092019-07-01 Generation and Characterization of Induced Pluripotent Stem Cells from Mononuclear Cells in Schizophrenic Patients Liu, Qing Du, Jiang Fan, Jinyu Li, Wenqiang Guo, Weiyun Feng, Huigen Lin, Juntang Cell J Original Article OBJECTIVE: Schizophrenia (SZ) is a mental disorder in which psychotic symptoms are the main problem. The pathogenesis of SZ is not fully understood, partly because of limitations in current disease models and technology. The development of induced pluripotent stem cell (iPSC) technology has opened up the possibility of elucidating disease mechanisms in neurodegenerative diseases. Here, we aimed to obtain iPSCs from peripheral blood mononuclear cells (PBMCs) of normal and schizophrenic individuals and analyze the inflammatory response in these iPSCs. MATERIALS AND METHODS: In this experimental study, we isolated PBMCs from whole blood of healthy individuals and SZ patients and reprogrammed them into iPSCs by transfection of recombinant lentiviruses that contained Yamanaka factors (Oct4, Sox2, Klf4 and c-Myc). We calculated the numbers of iPSC clones and stained them with alkaline phosphatase (ALP), Nanog, SSEA4, Nestin, Vimentin, and AFP to confirm their efficiency and pluripotency. The iPSCs were analyzed by real-time quantitative polymerase chain reaction (qRT-PCR) for the expressions of inflammatory factors. RESULTS: iPSCs from schizophrenic patients (SZ-iPSCs) exhibited typical morphology and highly expressed pluripotent markers. These iPSCs retained their normal karyotype and differentiated in vitro to form embryoid bodies (EBs) that expressed markers of all 3 germ layers. However, iPSCs from the SZ-iPSCs group had a weak capacity to differentiate into ectoderm compared to the normal iPSCs (Con-iPSC). An elevated, stronger inflammatory response existed in iPSCs from schizophrenic individuals. CONCLUSION: We successfully obtained iPSCs from PBMCs of schizophrenic patients without genetic operation and analyzed the expressions of pluripotent markers and inflammatory factors between the Con-iPSC and SZ-iPSC groups. Taken together, our results may assist to explain the pathogenesis of SZ and develop new strategies for clinical diagnosis and treatment. Royan Institute 2019 2019-02-25 /pmc/articles/PMC6397609/ /pubmed/30825289 http://dx.doi.org/10.22074/cellj.2019.5871 Text en The Cell Journal (Yakhteh) is an open access journal which means the articles are freely available online for any individual author to download and use the providing address. http://creativecommons.org/licenses/by/3.0/ The journal is licensed under a Creative Commons Attribution-Non Commercial 3.0 Unported License which allows the author(s) to hold the copyright without restrictions that is permitting unrestricted use, distribution, and reproduction in any medium provided the original work is properly cited. |
spellingShingle | Original Article Liu, Qing Du, Jiang Fan, Jinyu Li, Wenqiang Guo, Weiyun Feng, Huigen Lin, Juntang Generation and Characterization of Induced Pluripotent Stem Cells from Mononuclear Cells in Schizophrenic Patients |
title | Generation and Characterization of Induced Pluripotent
Stem Cells from Mononuclear Cells in
Schizophrenic Patients |
title_full | Generation and Characterization of Induced Pluripotent
Stem Cells from Mononuclear Cells in
Schizophrenic Patients |
title_fullStr | Generation and Characterization of Induced Pluripotent
Stem Cells from Mononuclear Cells in
Schizophrenic Patients |
title_full_unstemmed | Generation and Characterization of Induced Pluripotent
Stem Cells from Mononuclear Cells in
Schizophrenic Patients |
title_short | Generation and Characterization of Induced Pluripotent
Stem Cells from Mononuclear Cells in
Schizophrenic Patients |
title_sort | generation and characterization of induced pluripotent
stem cells from mononuclear cells in
schizophrenic patients |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6397609/ https://www.ncbi.nlm.nih.gov/pubmed/30825289 http://dx.doi.org/10.22074/cellj.2019.5871 |
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