目的：研究人胚胎植入前囊胚滋养外胚层细胞基因的空间表达。方法：辅助生殖来源人受精后第6天Gardner评分5AA的囊胚，显微镜下微吸管机械分离滋养外胚层细胞，对囊胚滋养外胚层极滋养层细胞和壁滋养层细胞采用单细胞测序检测，选取经t检验计算所得P＜0.05且基因表达差异≥2倍的差异表达基因，采用生物信息学软件对筛选的差异基因进行无监督层次聚类分析和基因本体（gene ontology，GO）功能分类分析，对差异表达基因进行注释和生物功能富集，并用全基因组对差异基因功能进一步注释。结果：对2枚受精后第6天的5AA囊胚滋养外胚层8个极滋养层细胞和8个壁滋养层细胞测序，发现极滋养层细胞306个基因上调，壁滋养层细胞75个基因上调。无监督聚类分析结果显示外胚层细胞分成极滋养层细胞和壁滋养层细胞两群，属于两种不同类型和生物功能的细胞群。差异基因功能注释显示，极滋养层细胞上调基因GO生物学功能主要是转录、能量代谢、蛋白合成、转运、氧化应激、离子转运、蛋白合成及运输、细胞周期调节、肌动蛋白增长等，主要参与泛素介导的蛋白水解、氧化磷酸化、Wnt信号通路、雌雄激素代谢等信号通路；壁滋养层细胞上调基因GO生物学功能主要是蛋白分解代谢、细胞周期停滞、细胞凋亡、激活丝裂原活化蛋白激酶（mitogenactivated protein kinase，MAPK）信号通路、碳水化合物运输、突触负调节、细胞生长、钙通道激活、正向B 细胞分化、T细胞凋亡等，主要参与B细胞受体、T细胞受体、白细胞跨内皮移植、血管内皮生长因子（vascular endothelial growth factor，VEGF）表达、间隙连接、促性腺激素释放激素（gonadotropin-releasing hormone，GnRH）分泌、细胞凋亡等信号通路。结论：从空间维度揭示囊胚极/ 壁滋养层细胞的基因表达，显示胚胎植入前囊胚极/壁滋养外胚层相互协调，精细调节囊胚孵化和胚胎植入过程，进一步数据分析有望寻找到调控胚胎植入的内源性特异分子。

Objective： To study the spatial expression of trophectoderm cells in human embryonic preantral blastocysts. Methods: The study used Gardner score 5AA blastocysts harvested on day 6 after fertilization from assisted reproductive technology. Microcapsules were used to separate trophectoderm cells from the epidermal cells. Single-cell sequencing was performed. P ＜ 0.05 was calculated by unpaired t test, and the difference was 2 times. Here we determined, for the first time, global gene expression patterns in the polar/mural trophectoderm isolated from human blastocysts. Unsupervised hierarchical clustering analysis and gene ontology (GO) functional classification were performed using bioinformatics software. Differentially expressed genes were annotated by the Database for Annotation, Visualization and Integrated Discovery. Functions of differentially expressed genes were further annotated using encyclopedia of genes and genomes. Results: The results showed that there were up to 306 genes in the trophoblast cells and up to 75 genes in the trophoblast cells. Unsupervised cluster analysis of polar trophoblast cells and mural trophoblast cells were divided into two groups, belonging to different types and biological functions. Differences in gene function indicated that the biological functions of GO gene uptake genes were mainly transcription, energy metabolism, protein synthesis, transport, oxidative stress, ion transport, protein synthesis and transport, cell cycle regulation, actin growth, etc. They were mainly involved in ubiquitin-mediated protein hydrolysis, oxidative phosphorylation, Wnt signaling pathway, estrogen androgen metabolism and other signal pathways; wall trophoblast cells up-regulated gene GO biological function, which was mainly proteolytic metabolism, cell cycle arrest, apoptosis, activation of MAPK, carbohydrate transport, synaptic regulation, cell growth, calcium channel activation, positive B cell differentiation, T cell apoptosis and other biological functions, which were mainly involved in B cell receptor, T cell receptor, white blood cells crossendothelial transplantation, VEGF expression, gap connection, GnRH secretion, apoptosis and other signaling pathways. Conclusion: The gene expression of blastocysts trophectoderm is revealed from the spatial dimension, indicating that differentiation of polar and mural trophectoderm of blastocysts is accompanied by differences between the two cell lineages, and the polar and mural trophectoderms are coordinated with each other and the blastocyst hatching and embryo implantation processes are finely adjusted. Further data analysis is expected to find the endogenous molecular specificity of the regulation of embryo implantation.