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 multif...
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Wiley
2020-01-01
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Series: | Stem Cells International |
Online Access: | http://dx.doi.org/10.1155/2020/8897494 |
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author | Sijun Liu Yuying Zhao Xiaoying Su Chengcheng Zhou Peifen Yang Qiusan Lin Shijun Li Hanxu Tan Qi Wang Changjun Wang Qingguang Wu |
author_facet | Sijun Liu Yuying Zhao Xiaoying Su Chengcheng Zhou Peifen Yang Qiusan Lin Shijun Li Hanxu Tan Qi Wang Changjun Wang Qingguang Wu |
author_sort | Sijun Liu |
collection | DOAJ |
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 | Article |
id | doaj-art-f4063822a5f848e38788f0b5babc0334 |
institution | Kabale University |
issn | 1687-966X 1687-9678 |
language | English |
publishDate | 2020-01-01 |
publisher | Wiley |
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spelling | doaj-art-f4063822a5f848e38788f0b5babc03342025-02-03T05:52:58ZengWileyStem Cells International1687-966X1687-96782020-01-01202010.1155/2020/88974948897494Reconstruction of Alzheimer’s Disease Cell Model In Vitro via Extracted Peripheral Blood Molecular Cells from a Sporadic PatientSijun Liu0Yuying Zhao1Xiaoying Su2Chengcheng Zhou3Peifen Yang4Qiusan Lin5Shijun Li6Hanxu Tan7Qi Wang8Changjun Wang9Qingguang Wu10School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou 510006, ChinaInstitute of Clinical Pharmacology, Guangzhou University of Chinese Medicine, Guangzhou 510006, ChinaSchool of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou 510006, ChinaSchool of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou 510006, ChinaSchool of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou 510006, ChinaSchool of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou 510006, ChinaRadiology of Chinese PLA General Hospital, Beijing 100853, ChinaDongzhimen Hospital, Beijing University of Chinese Medicine, 100029, ChinaInstitute of Clinical Pharmacology, Guangzhou University of Chinese Medicine, Guangzhou 510006, ChinaGuangdong Geriatric Institute, Guangdong Provincial People’s Hospital, Guangzhou 510080, ChinaSchool of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou 510006, ChinaThe 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.http://dx.doi.org/10.1155/2020/8897494 |
spellingShingle | Sijun Liu Yuying Zhao Xiaoying Su Chengcheng Zhou Peifen Yang Qiusan Lin Shijun Li Hanxu Tan Qi Wang Changjun Wang Qingguang Wu Reconstruction of Alzheimer’s Disease Cell Model In Vitro via Extracted Peripheral Blood Molecular Cells from a Sporadic Patient Stem Cells International |
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 |
url | http://dx.doi.org/10.1155/2020/8897494 |
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