Yeh Hana, De Cruz Matthew A, You Yang, Ikezu Seiko, Ikezu Tsuneya
Graduate Program in Neuroscience, Boston University, Boston, United States.
Department of Pharmacology and Experimental Therapeutics, Boston University School of Medicine, Boston, United States.
Sci Rep. 2025 Jan 25;15(1):3207. doi: 10.1038/s41598-025-87543-1.
There are few in vitro models available to study microglial physiology in a homeostatic context. Recent approaches include the human induced pluripotent stem cell model, but these can be challenging for large-scale assays and may lead to batch variability. To advance our understanding of microglial biology while enabling scalability for high-throughput assays, we developed an inducible immortalized murine microglial cell line using a tetracycline expression system. The addition of doxycycline facilitates rapid cell proliferation, allowing for population expansion. Upon withdrawal of doxycycline, this monoclonal microglial cell line differentiates, resembling in vivo microglial physiology as demonstrated by the expression of microglial genes, innate immune responses, chemotaxis, and phagocytic abilities. We utilized live imaging and various molecular techniques to functionally characterize the clonal 2E11murine microglial cell line. Transcriptomic analysis showed that the 2E11 line exhibited characteristics of immature, proliferative microglia during doxycycline induction, and further differentiation led to a more homeostatic phenotype. Treatment with transforming growth factor-β modified the transcriptome of the 2E11 cell line, affecting cellular immune pathways. Our findings indicate that the 2E11 inducible immortalized cell line is a practical and convenient tool for studying microglial biology in vitro.
在稳态环境下，用于研究小胶质细胞生理学的体外模型很少。最近的方法包括人类诱导多能干细胞模型，但这些模型对于大规模检测可能具有挑战性，并且可能导致批次差异。为了增进我们对小胶质细胞生物学的理解，同时实现高通量检测的可扩展性，我们使用四环素表达系统开发了一种可诱导的永生化小鼠小胶质细胞系。加入强力霉素可促进细胞快速增殖，从而实现细胞数量的扩增。去除强力霉素后，这种单克隆小胶质细胞系会发生分化，通过小胶质细胞基因的表达、先天免疫反应、趋化性和吞噬能力等方面表现出与体内小胶质细胞生理学相似的特征。我们利用实时成像和各种分子技术对克隆的2E11小鼠小胶质细胞系进行功能表征。转录组分析表明，在强力霉素诱导期间，2E11细胞系表现出未成熟、增殖性小胶质细胞的特征，进一步分化则导致更接近稳态的表型。用转化生长因子-β处理可改变2E11细胞系的转录组，影响细胞免疫途径。我们的研究结果表明，2E11可诱导永生化细胞系是体外研究小胶质细胞生物学的一种实用且便捷的工具。
