图1:mhCOs中小胶质样细胞的特征。生成mhCOs的原理图。将10%的PU.1感染hESCs与90%的亲本HES3 hESCs混合，分别在第2天和第18天进行PU.1启动和完全诱导。b对照hCOs和mhCOs(30天和70天)小胶质细胞相关基因的表达。在第30天测量相对于对照类器官的基因表达，归一化至β-肌动蛋白。数据代表平均值±SEM (n = 5，三个独立批次)。c左，IBA1免疫染色显示切片mhcos在第30天和第70天产生小胶质细胞样细胞。对照hCOs中未发现IBA1+细胞。右，不同时间点mhCOs中IBA1 +小胶质细胞样细胞的Sholl分析。数据代表平均值±SEM (n = 5个类器官，来自两个hESCs系的三个独立分化重复)。 d and e Immunostaining of mhCOs at day 70 and isolated microglia co-cultured with neurons (2D) for IBA1 and CSF1R (d) TMEM119 and P2RY12 (e). Representative images were shown (n = 5, from two independent batches). f Top, co-expression of PU.1 and IBA1 in hCOs and mhCOs at day 30 and 70. Bottom, quantification of Pu.1-derived IBA1 microglia-like cells. Data represent the mean ± SEM (n = 5 organoids from three independent differentiation replicates of two hESCs lines). Bottom, Pearson’s correlation coefficient of IBA1 with PU.1 in mhCOs at days 30 and 70. g Top, co-immunostaining for Ki67 and IBA1 in mhCOs at day 70. Bottom, quantification of proliferating IBA1 microglia-like cells. Data represent the mean ± SEM (n = 5 organoids from three independent differentiation replicates of two hESC lines). h Left, high-resolution imaging showed microglia isolated from mhCOs at day 90 and co-cultured D90 cortical neurons for 3 days contained inclusions of PSD95. Right, quantification of PSD95 particles in IBA1+ microglia-like cells. Data represent the mean ± SEM (n = 8, from three independent differentiation replicates of hESCs lines). The scale bar represents 50 μm in c–g and 20 μm in h. Credit: DOI: 10.1038/s41467-022-28043-y
