Cargando…
Reconstruction of Alzheimer's Disease Cell Model In Vitro via Extracted Peripheral Blood Molecular Cells from a Sporadic Patient
The establishment of human-induced pluripotent stem cell (iPSC) models from sporadic Alzheimer's disease (sAD) patients is necessary and could potentially benefit research into disease etiology and therapeutic strategies. However, the development of sAD iPSC models is still limited due to the m...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7762651/ https://www.ncbi.nlm.nih.gov/pubmed/33381193 http://dx.doi.org/10.1155/2020/8897494 |
_version_ | 1783627856444653568 |
---|---|
author | Liu, Sijun Zhao, Yuying Su, Xiaoying Zhou, Chengcheng Yang, Peifen Lin, Qiusan Li, Shijun Tan, Hanxu Wang, Qi Wang, Changjun Wu, Qingguang |
author_facet | Liu, Sijun Zhao, Yuying Su, Xiaoying Zhou, Chengcheng Yang, Peifen Lin, Qiusan Li, Shijun Tan, Hanxu Wang, Qi Wang, Changjun Wu, Qingguang |
author_sort | Liu, Sijun |
collection | PubMed |
description | The establishment of human-induced pluripotent stem cell (iPSC) models from sporadic Alzheimer's disease (sAD) patients is necessary and could potentially benefit research into disease etiology and therapeutic strategies. However, the development of sAD iPSC models is still limited due to the multifactorial nature of the disease. Here, we extracted peripheral blood mononuclear cells (PBMCs) from a patient with sAD and induced them into iPSC by introducing the Sendai virus expressing Oct3/4, Sox2, c-Myc, and Klf4, which were subsequently induced into neural cells to build the cell model of AD. Using alkaline phosphatase staining, immunofluorescence staining, karyotype analysis, reverse transcription-polymerase chain reaction (RT-PCR), and teratoma formation in vitro, we demonstrated that the iPSC derived from PMBCs (PBMC-iPSC) had a normal karyotype and potential to differentiate into three embryonic layers. Immunofluorescence staining and quantitative real-time polymerase chain reaction (qPCR) suggested that PBMC-iPSCs were successfully differentiated into neural cells. Detection of beta-amyloid protein oligomer (AβO), beta-amyloid protein 1-40 (Aβ 1-40), and beta-amyloid protein 1-42 (Aβ 1-42) indicated that the AD cell model was satisfactorily constructed in vitro. In conclusion, this study has successfully generated an AD cell model with pathological features of beta-amyloid peptide deposition using PBMC from a patient with sAD. |
format | Online Article Text |
id | pubmed-7762651 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Hindawi |
record_format | MEDLINE/PubMed |
spelling | pubmed-77626512020-12-29 Reconstruction of Alzheimer's Disease Cell Model In Vitro via Extracted Peripheral Blood Molecular Cells from a Sporadic Patient Liu, Sijun Zhao, Yuying Su, Xiaoying Zhou, Chengcheng Yang, Peifen Lin, Qiusan Li, Shijun Tan, Hanxu Wang, Qi Wang, Changjun Wu, Qingguang Stem Cells Int Research Article The establishment of human-induced pluripotent stem cell (iPSC) models from sporadic Alzheimer's disease (sAD) patients is necessary and could potentially benefit research into disease etiology and therapeutic strategies. However, the development of sAD iPSC models is still limited due to the multifactorial nature of the disease. Here, we extracted peripheral blood mononuclear cells (PBMCs) from a patient with sAD and induced them into iPSC by introducing the Sendai virus expressing Oct3/4, Sox2, c-Myc, and Klf4, which were subsequently induced into neural cells to build the cell model of AD. Using alkaline phosphatase staining, immunofluorescence staining, karyotype analysis, reverse transcription-polymerase chain reaction (RT-PCR), and teratoma formation in vitro, we demonstrated that the iPSC derived from PMBCs (PBMC-iPSC) had a normal karyotype and potential to differentiate into three embryonic layers. Immunofluorescence staining and quantitative real-time polymerase chain reaction (qPCR) suggested that PBMC-iPSCs were successfully differentiated into neural cells. Detection of beta-amyloid protein oligomer (AβO), beta-amyloid protein 1-40 (Aβ 1-40), and beta-amyloid protein 1-42 (Aβ 1-42) indicated that the AD cell model was satisfactorily constructed in vitro. In conclusion, this study has successfully generated an AD cell model with pathological features of beta-amyloid peptide deposition using PBMC from a patient with sAD. Hindawi 2020-12-18 /pmc/articles/PMC7762651/ /pubmed/33381193 http://dx.doi.org/10.1155/2020/8897494 Text en Copyright © 2020 Sijun Liu 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 Liu, Sijun Zhao, Yuying Su, Xiaoying Zhou, Chengcheng Yang, Peifen Lin, Qiusan Li, Shijun Tan, Hanxu Wang, Qi Wang, Changjun Wu, Qingguang Reconstruction of Alzheimer's Disease Cell Model In Vitro via Extracted Peripheral Blood Molecular Cells from a Sporadic Patient |
title | Reconstruction of Alzheimer's Disease Cell Model In Vitro via Extracted Peripheral Blood Molecular Cells from a Sporadic Patient |
title_full | Reconstruction of Alzheimer's Disease Cell Model In Vitro via Extracted Peripheral Blood Molecular Cells from a Sporadic Patient |
title_fullStr | Reconstruction of Alzheimer's Disease Cell Model In Vitro via Extracted Peripheral Blood Molecular Cells from a Sporadic Patient |
title_full_unstemmed | Reconstruction of Alzheimer's Disease Cell Model In Vitro via Extracted Peripheral Blood Molecular Cells from a Sporadic Patient |
title_short | Reconstruction of Alzheimer's Disease Cell Model In Vitro via Extracted Peripheral Blood Molecular Cells from a Sporadic Patient |
title_sort | reconstruction of alzheimer's disease cell model in vitro via extracted peripheral blood molecular cells from a sporadic patient |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7762651/ https://www.ncbi.nlm.nih.gov/pubmed/33381193 http://dx.doi.org/10.1155/2020/8897494 |
work_keys_str_mv | AT liusijun reconstructionofalzheimersdiseasecellmodelinvitroviaextractedperipheralbloodmolecularcellsfromasporadicpatient AT zhaoyuying reconstructionofalzheimersdiseasecellmodelinvitroviaextractedperipheralbloodmolecularcellsfromasporadicpatient AT suxiaoying reconstructionofalzheimersdiseasecellmodelinvitroviaextractedperipheralbloodmolecularcellsfromasporadicpatient AT zhouchengcheng reconstructionofalzheimersdiseasecellmodelinvitroviaextractedperipheralbloodmolecularcellsfromasporadicpatient AT yangpeifen reconstructionofalzheimersdiseasecellmodelinvitroviaextractedperipheralbloodmolecularcellsfromasporadicpatient AT linqiusan reconstructionofalzheimersdiseasecellmodelinvitroviaextractedperipheralbloodmolecularcellsfromasporadicpatient AT lishijun reconstructionofalzheimersdiseasecellmodelinvitroviaextractedperipheralbloodmolecularcellsfromasporadicpatient AT tanhanxu reconstructionofalzheimersdiseasecellmodelinvitroviaextractedperipheralbloodmolecularcellsfromasporadicpatient AT wangqi reconstructionofalzheimersdiseasecellmodelinvitroviaextractedperipheralbloodmolecularcellsfromasporadicpatient AT wangchangjun reconstructionofalzheimersdiseasecellmodelinvitroviaextractedperipheralbloodmolecularcellsfromasporadicpatient AT wuqingguang reconstructionofalzheimersdiseasecellmodelinvitroviaextractedperipheralbloodmolecularcellsfromasporadicpatient |