Efficient conversion of human induced pluripotent stem cells into microglia by defined transcription factors.
Efficient conversion of human induced pluripotent stem cells into microglia by defined transcription factors.
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DOI：
10.1016/j.stemcr.2021.03.010
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发表时间：
2021-05-11
影响因子：
5.9
通讯作者：
Wong YH
中科院分区：
文献类型：
--
作者：
Chen SW;Hung YS;Fuh JL;Chen NJ;Chu YS;Chen SC;Fann MJ;Wong YH
小胶质细胞是中枢神经系统的免疫细胞，在脑生理和病理中发挥着重要作用。我们报道了一种新的方法，通过强制表达SPI1和CEBPA，在10天内将人诱导多能干细胞(HiPSCs)分化为小胶质细胞(IMG)。RT-qPCR、免疫染色和流式细胞仪分析证实IMG细胞的主要小胶质细胞标志物的高水平表达和纯度。全转录组分析表明，这些IMG类似于人胎儿/成人小胶质细胞，而不是人类单核细胞。此外，这些IMGs还具有适当的生理功能，包括各种炎症反应、ADP/ATP诱导的迁移和吞噬能力。当IMGs与HiPSC来源的神经元共同培养时，IMGs会做出反应并向受损的神经元迁移。这项研究建立了一种将HiPSCs快速转化为功能性IMG的方案，这将有助于使用不同的疾病模型对人类小胶质细胞进行功能研究，并有助于药物的发现。通过强制表达SPI1和CEBPA从IPSCs中高效生成人IMG IMG的转录组特征与人原代小胶质细胞相似，IMG细胞具有适当的生理功能。报道了一种新的方法，通过强制表达SPI1和CEBPA，在10天内从诱导的多能干细胞中产生人小胶质细胞(IMG)。这些IMG具有适当的生理功能，可以与HiPSC来源的神经元共培养，从而可以在体外研究正常和疾病条件下神经元与小胶质细胞的相互作用。
Microglia, the immune cells of the central nervous system, play critical roles in brain physiology and pathology. We report a novel approach that produces, within 10 days, the differentiation of human induced pluripotent stem cells (hiPSCs) into microglia (iMG) by forced expression of both SPI1 and CEBPA. High-level expression of the main microglial markers and the purity of the iMG cells were confirmed by RT-qPCR, immunostaining, and flow cytometry analyses. Whole-transcriptome analysis demonstrated that these iMGs resemble human fetal/adult microglia but not human monocytes. Moreover, these iMGs exhibited appropriate physiological functions, including various inflammatory responses, ADP/ATP-evoked migration, and phagocytic ability. When co-cultured with hiPSC-derived neurons, the iMGs respond and migrate toward injured neurons. This study has established a protocol for the rapid conversion of hiPSCs into functional iMGs, which should facilitate functional studies of human microglia using different disease models and also help with drug discovery. Efficient generation of human iMGs from iPSCs by forced expression of SPI1 and CEBPA The transcriptome profile of iMGs resembles that of human primary microglia The iMG cells possess appropriate physiological functioning An iN-iMG co-culture model is established for studying neuron-microglia interactions Chen et al. report a novel approach that produces human microglia (iMG) from induced pluripotent stem cells within 10 days by forced expression of SPI1 and CEBPA together. These iMGs exhibit appropriate physiological functions and can be co-cultured with hiPSC-derived neurons, allowing the study of in vitro neuron-microglia interactions under both normal and disease conditions.
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影响因子：
5.1
作者：
Fu, Ruying;Shen, Qingyu;Xu, Pengfei;Luo, Jin Jun;Tang, Yamei
通讯作者：
Tang, Yamei
影响因子：
25
作者：
Pandya H;Shen MJ;Ichikawa DM;Sedlock AB;Choi Y;Johnson KR;Kim G;Brown MA;Elkahloun AG;Maric D;Sweeney CL;Gossa S;Malech HL;McGavern DB;Park JK
通讯作者：
Park JK
影响因子：
5.9
作者：
Douvaras P;Sun B;Wang M;Kruglikov I;Lallos G;Zimmer M;Terrenoire C;Zhang B;Gandy S;Schadt E;Freytes DO;Noggle S;Fossati V
通讯作者：
Fossati V
影响因子：
56.9
作者：
Nimmerjahn, A;Kirchhoff, F;Helmchen, F
通讯作者：
Helmchen, F
影响因子：
25
作者：
Kierdorf, Katrin;Erny, Daniel;Prinz, Marco
通讯作者：
Prinz, Marco