Reconstruction of Alzheimer's Disease Cell Model In Vitro via Extracted Peripheral Blood Molecular Cells from a Sporadic Patient

Sijun Liu; Yuying Zhao; Xiaoying Su; Chengcheng Zhou; Peifen Yang; Qiusan Lin; Shijun Li; Hanxu Tan; Qi Wang; Changjun Wang; Qingguang Wu

doi:10.1155/2020/8897494

Reconstruction of Alzheimer's Disease Cell Model In Vitro via Extracted Peripheral Blood Molecular Cells from a Sporadic Patient

Stem Cells Int. 2020 Dec 18:2020:8897494. doi: 10.1155/2020/8897494. eCollection 2020.

Authors

Sijun Liu^{1

2}, Yuying Zhao³, Xiaoying Su¹, Chengcheng Zhou¹, Peifen Yang¹, Qiusan Lin¹, Shijun Li^{4

5}, Hanxu Tan⁶, Qi Wang³, Changjun Wang⁷, Qingguang Wu¹

Affiliations

¹ School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou 510006, China.
² Beth Israel Deaconess Medical Center, Department of Surgery, Harvard Medical School, 02120, USA.
³ Institute of Clinical Pharmacology, Guangzhou University of Chinese Medicine, Guangzhou 510006, China.
⁴ Radiology of Chinese PLA General Hospital, Beijing 100853, China.
⁵ Martinos Center of Harvard Medical School, MA 02129, USA.
⁶ Dongzhimen Hospital, Beijing University of Chinese Medicine, 100029, China.
⁷ Guangdong Geriatric Institute, Guangdong Provincial People's Hospital, Guangzhou 510080, China.

Abstract

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.