Cell-surface proteomics identifies lineage-specific markers of embryo-derived stem cells.

The advent of reprogramming and its impact on stem cell biology has renewed interest in lineage restriction in mammalian embryos, the source of embryonic (ES), epiblast (EpiSC), trophoblast (TS), and extraembryonic endoderm (XEN) stem cell lineages. Isolation of specific cell types during stem cell differentiation and reprogramming, and also directly from embryos, is a major technical challenge because few cell-surface proteins are known that can distinguish each cell type. We provide a large-scale proteomic resource of cell-surface proteins for the four embryo-derived stem cell lines. We validated 27 antibodies against lineage-specific cell-surface markers, which enabled investigation of specific cell populations during ES-EpiSC reprogramming and ES-to-XEN differentiation. Identified markers also allowed prospective isolation and characterization of viable lineage progenitors from blastocysts by flow cytometry. These results provide a comprehensive stem cell proteomic resource and enable new approaches to interrogate the mechanisms that regulate cell fate specification. ► Cell-surface proteome for four embryo-derived stem cell lineages ► Comprehensive resource of signaling, adhesion and migration proteins ► Enables isolation of specific cell types during differentiation and reprogramming ► Allows prospective isolation of lineage progenitors directly from blastocysts Rugg-Gunn et al. provide a large-scale proteomic resource of cell-surface proteins for four embryo-derived stem cell lines: ES, EpiSC, TS, and XEN. Antibodies against lineage-specific cell-surface markers allowed investigation of specific cell populations during differentiation and reprogramming and also prospective isolation of viable lineage progenitors from blastocysts by flow cytometry.

重编程的出现及其对干细胞生物学的影响，使人们对哺乳动物胚胎中的谱系限制重新产生了兴趣，哺乳动物胚胎是胚胎（ES）、上胚层（EpiSC）、滋养层（TS）和胚外内胚层（XEN）干细胞谱系的来源。在干细胞分化和重编程过程中，以及直接从胚胎中分离特定细胞类型是一项重大的技术挑战，因为已知能够区分每种细胞类型的细胞表面蛋白很少。我们为四种胚胎来源的干细胞系提供了大规模的细胞表面蛋白质组学资源。我们验证了27种针对谱系特异性细胞表面标志物的抗体，这使得能够在ES - EpiSC重编程和ES到XEN分化过程中对特定细胞群体进行研究。所鉴定的标志物还允许通过流式细胞术从囊胚中前瞻性地分离和鉴定有活力的谱系祖细胞。这些结果提供了一个全面的干细胞蛋白质组学资源，并使人们能够采用新的方法来探究调节细胞命运特化的机制。 ► 四种胚胎来源干细胞谱系的细胞表面蛋白质组 ► 信号传导、黏附和迁移蛋白质的综合资源 ► 能够在分化和重编程过程中分离特定细胞类型 ► 允许直接从囊胚中前瞻性地分离谱系祖细胞 拉格 - 冈恩等人（Rugg - Gunn et al.）为四种胚胎来源的干细胞系（ES、EpiSC、TS和XEN）提供了大规模的细胞表面蛋白质组学资源。针对谱系特异性细胞表面标志物的抗体使得能够在分化和重编程过程中对特定细胞群体进行研究，并且还能够通过流式细胞术从囊胚中前瞻性地分离有活力的谱系祖细胞。