Gonul Ceren Perihan, Kiser Cagla, Yaka Emis Cansu, Oz Didem, Hunerli Duygu, Yerlikaya Deniz, Olcum Melis, Keskinoglu Pembe, Yener Gorsev, Genc Sermin
Izmir Biomedicine and Genome Center, Dokuz Eylul University Health Campus, Izmir, Türkiye; Izmir International Biomedicine and Genome Institute, Dokuz Eylul University, Izmir, Türkiye.
Department of Neuroscience, Institute of Health Sciences, Dokuz Eylul University, Izmir, Türkiye; Department of Neurology, Izmir City Hospital, Izmir, Türkiye.
Mol Cell Neurosci. 2025 Mar;132:103990. doi: 10.1016/j.mcn.2024.103990. Epub 2024 Dec 27.
Alzheimer's disease (AD) is a neurodegenerative disorder that is characterized by the accumulation of amyloid plaques, phosphorylated tau tangles and microglia toxicity, resulting in neuronal death and cognitive decline. Since microglia are recognized as one of the key players in the disease, it is crucial to understand how microglia operate in disease conditions and incorporate them into models. The studies on human microglia functions are thought to reflect the post-symptomatic stage of the disease. Recently developed methods involve induced microglia-like cells (iMGs) generated from patients' blood monocytes or induced pluripotent stem cells (iPSCs) as an alternative to studying the microglia cells in vitro. In this research, we aimed to investigate the phenotype and inflammatory responses of iMGs from AD patients. Monocytes derived from blood using density gradient centrifugation were differentiated into iMGs using a cytokine cocktail, including granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-34 (IL-34). After differentiation, cells were assessed by morphological analysis and a microglia surface marker, TMEM119. We used stimulants, lipopolysaccharide (LPS) and beta-amyloid, to examine iMGs' functions. Results showed that iMGs derived from AD patients exhibited increased secretion of pro-inflammatory cytokines upon LPS stimulation. Furthermore, their phagocytic ability was also heightened in stimulated and unstimulated conditions, with cells derived from patients showing increased phagocytic activity compared to healthy controls. Overall, these findings suggest that iMGs derived from patients using the direct conversion method possess characteristics of human microglia, making them an easy and promising model for studying microglia function in AD.
阿尔茨海默病（AD）是一种神经退行性疾病，其特征在于淀粉样斑块的积累、磷酸化tau缠结和小胶质细胞毒性，导致神经元死亡和认知能力下降。由于小胶质细胞被认为是该疾病的关键因素之一，了解小胶质细胞在疾病状态下的运作方式并将其纳入模型至关重要。对人类小胶质细胞功能的研究被认为反映了该疾病的症状后阶段。最近开发的方法涉及从患者血液单核细胞或诱导多能干细胞（iPSC）生成诱导性小胶质细胞样细胞（iMG），作为体外研究小胶质细胞的替代方法。在本研究中，我们旨在研究AD患者来源的iMG的表型和炎症反应。使用密度梯度离心法从血液中分离出的单核细胞，通过细胞因子鸡尾酒（包括粒细胞-巨噬细胞集落刺激因子（GM-CSF）和白细胞介素-34（IL-34））分化为iMG。分化后，通过形态分析和小胶质细胞表面标志物TMEM119对细胞进行评估。我们使用刺激物脂多糖（LPS）和β-淀粉样蛋白来检测iMG的功能。结果表明，AD患者来源的iMG在LPS刺激下促炎细胞因子的分泌增加。此外，在刺激和未刺激条件下，它们的吞噬能力也有所增强，与健康对照相比，患者来源的细胞吞噬活性增加。总体而言，这些发现表明，使用直接转化方法从患者获得的iMG具有人类小胶质细胞的特征，使其成为研究AD中小胶质细胞功能的一种简便且有前景的模型。
