Transcriptional changes during astrocyte maturation in vivo and in culture

Analysis of gene expression during astrocyte differentiation and maturation in vivo and in vitro in mouse, and investigation of transcription factors and extrinsic conditions influencing maturation In vivo astrocytes were purified from mice using anti-ACSA2 MACS at indicated ages. In vitro astrocytes were differentiated from cultured neural stem/progenitor cells (cultured in EGF/FGF2 medium) using BMP4 for 14 days (without EGF/FGF2). Where indicated, astrocytes in vitro were infected with transgenes and gene expression induced from day 7 to 14, or astrocytes differentiated for 7 days in BMP4 in normal 2D cultures or in 3D cultures were matured for 7 days in basal medium or FGF2 containing medium. Cells were analysed by bulk RNA-Seq.小鼠体内外星形胶质细胞（astrocyte）分化与成熟过程中的基因表达分析，以及对影响其成熟的转录因子与外源调控条件的探究。研究中，体内星形胶质细胞按照指定日龄，采用抗ACSA2磁激活细胞分选（MACS）技术从小鼠体内分离纯化；体外星形胶质细胞则由培养于EGF/FGF2培养基中的神经干细胞/前体细胞诱导分化而来：使用BMP4诱导分化14天，期间不添加EGF/FGF2。若有指定实验分组，体外星形胶质细胞可通过转染外源基因，并于第7至14天诱导其基因表达；或是将经BMP4诱导分化7天的星形胶质细胞，分别置于常规二维（2D）培养体系或三维（3D）培养体系中，再以基础培养基或添加FGF2的培养基继续成熟培养7天。所有细胞样本均通过批量RNA测序（bulk RNA-Seq）进行转录组分析。