Single cell RNA-seq of ten-thousand epithelium cells from the mammary glands of mice in puberty, estrus and diestrus phases

Epithelial cells were isolated by FACS from the mammary glands of pubescent (5 week old), estrus adult (10 week old) and diestrus adult (10 week old) female mice. Freshly sorted cells were submitted to a 10X Genomics Chromium System for single cell capture. cDNA synthesis and library preparation was done according to the protocol supplied by the manufacturer. Sequencing was carried out on an Illumina NextSeq500 sequencer using parameters recommended by 10X Genomics. Transcriptional profiling was completed for 5387 total epithelial cells from pubescent (5 week old) mice, 2729 total epithelial cells from adult (10 week old) mice in the estrus phase, and 2439 total epithelial cells from adult (10 week old) mice in the diestrus phase.研究人员通过荧光激活细胞分选术（Fluorescence-Activated Cell Sorting, FACS），分别从青春期（5周龄）、发情期成年（10周龄）及间情期成年（10周龄）雌性小鼠的乳腺组织中分离上皮细胞。将新鲜分选获得的细胞投入10X Genomics Chromium系统完成单细胞捕获，随后依照厂商提供的实验流程开展互补DNA（complementary DNA, cDNA）合成与文库构建工作。测序实验在Illumina NextSeq500测序仪上进行，测序参数参照10X Genomics官方推荐方案设置。最终完成了三类样本的转录组谱分析（transcriptional profiling），共获得5387个青春期（5周龄）小鼠乳腺上皮细胞、2729个发情期成年（10周龄）小鼠乳腺上皮细胞，以及2439个间情期成年（10周龄）小鼠乳腺上皮细胞的相关测序数据。