Liu Sijun, Zhao Yuying, Su Xiaoying, Zhou Chengcheng, Yang Peifen, Lin Qiusan, Li Shijun, Tan Hanxu, Wang Qi, Wang Changjun, Wu Qingguang
School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou 510006, China.
Beth Israel Deaconess Medical Center, Department of Surgery, Harvard Medical School, 02120, USA.
Stem Cells Int. 2020 Dec 18;2020:8897494. doi: 10.1155/2020/8897494. eCollection 2020.
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 , 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 (AO), beta-amyloid protein 1-40 (A 1-40), and beta-amyloid protein 1-42 (A 1-42) indicated that the AD cell model was satisfactorily constructed . 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.
从散发性阿尔茨海默病(sAD)患者中建立人诱导多能干细胞(iPSC)模型很有必要,可能有助于疾病病因和治疗策略的研究。然而,由于该疾病的多因素性质,sAD iPSC模型的发展仍然有限。在此,我们从一名sAD患者中提取外周血单个核细胞(PBMC),通过导入表达Oct3/4、Sox2、c-Myc和Klf4的仙台病毒将其诱导为iPSC,随后将这些iPSC诱导为神经细胞以构建AD细胞模型。通过碱性磷酸酶染色、免疫荧光染色、核型分析、逆转录-聚合酶链反应(RT-PCR)和畸胎瘤形成实验,我们证明源自PBMC的iPSC(PBMC-iPSC)具有正常核型且有分化为三个胚层的潜力。免疫荧光染色和定量实时聚合酶链反应(qPCR)表明PBMC-iPSC成功分化为神经细胞。β-淀粉样蛋白寡聚体(AO)、β-淀粉样蛋白1-40(A 1-40)和β-淀粉样蛋白1-42(A 1-42)的检测表明成功构建了AD细胞模型。总之,本研究利用一名sAD患者的PBMC成功构建了具有β-淀粉样肽沉积病理特征的AD细胞模型。