Banerjee Poulomi, Paza Evdokia, Perkins Emma M, James Owen G, Kenkhuis Boyd, Lloyd Amy F, Burr Karen, Story David, Yusuf Dilmurat, He Xin, Backofen Rolf, Dando Owen, Chandran Siddharthan, Priller Josef
Centre for Clinical Brain Sciences, University of Edinburgh, Edinburgh, UK; UK Dementia Research Institute at University of Edinburgh, Edinburgh, UK.
Centre for Clinical Brain Sciences, University of Edinburgh, Edinburgh, UK; UK Dementia Research Institute at University of Edinburgh, Edinburgh, UK; Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands.
Stem Cell Res. 2020 Dec;49:102046. doi: 10.1016/j.scr.2020.102046. Epub 2020 Oct 14.
Microglia are resident tissue macrophages of the central nervous system (CNS) that arise from erythromyeloid progenitors during embryonic development. They play essential roles in CNS development, homeostasis and response to disease. Since microglia are difficult to procure from the human brain, several protocols have been developed to generate microglia-like cells from human induced pluripotent stem cells (hiPSCs). However, some concerns remain over the purity and quality of in vitro generated microglia. Here, we describe a new protocol that does not require co-culture with neural cells and yields cultures of 100% P2Y 95% TMEM119 ramified human microglia-like cells (hiPSC-MG). In the presence of neural precursor cell-conditioned media, hiPSC-MG expressed high levels of human microglia signature genes, including SALL1, CSF1R, P2RY12, TMEM119, TREM2, HEXB and SIGLEC11, as revealed by whole-transcriptome analysis. Stimulation of hiPSC-MG with lipopolysaccharide resulted in downregulation of P2Y expression, induction of IL1B mRNA expression and increase in cell capacitance. HiPSC-MG were phagocytically active and maintained their cell identity after transplantation into murine brain slices and human brain spheroids. Together, our new protocol for the generation of microglia-like cells from human iPSCs will facilitate the study of human microglial function in health and disease.
小胶质细胞是中枢神经系统（CNS）中的常驻组织巨噬细胞，在胚胎发育过程中由红系髓系祖细胞产生。它们在中枢神经系统发育、稳态维持和疾病反应中发挥着重要作用。由于从小鼠大脑中获取小胶质细胞较为困难，因此已经开发了几种方案来从人诱导多能干细胞（hiPSC）中生成小胶质细胞样细胞。然而，对于体外生成的小胶质细胞的纯度和质量仍存在一些担忧。在这里，我们描述了一种新的方案，该方案不需要与神经细胞共培养，并且能够产生100% P2Y、95% TMEM119呈分支状的人小胶质细胞样细胞（hiPSC-MG）培养物。全转录组分析显示，在神经前体细胞条件培养基存在的情况下，hiPSC-MG表达高水平的人小胶质细胞特征基因，包括SALL1、CSF1R、P2RY12、TMEM119、TREM2、HEXB和SIGLEC11。用脂多糖刺激hiPSC-MG会导致P2Y表达下调、IL1B mRNA表达诱导以及细胞电容增加。hiPSC-MG具有吞噬活性，并且在移植到小鼠脑片和人脑类器官后仍保持其细胞身份。总之，我们从人诱导多能干细胞生成小胶质细胞样细胞的新方案将有助于研究健康和疾病状态下的人小胶质细胞功能。
