Institute of Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California, USA.
Nat Biotechnol. 2012 May 27;30(6):531-42. doi: 10.1038/nbt.2239.
To identify early populations of committed progenitors derived from human embryonic stem cells (hESCs), we screened self-renewing, BMP4-treated and retinoic acid-treated cultures with >400 antibodies recognizing cell-surface antigens. Sorting of >30 subpopulations followed by transcriptional analysis of developmental genes identified four distinct candidate progenitor groups. Subsets detected in self-renewing cultures, including CXCR4(+) cells, expressed primitive endoderm genes. Expression of Cxcr4 in primitive endoderm was confirmed in visceral endoderm of mouse embryos. BMP4-induced progenitors exhibited gene signatures of mesoderm, trophoblast and vascular endothelium, suggesting correspondence to gastrulation-stage primitive streak, chorion and allantois precursors, respectively. Functional studies in vitro and in vivo confirmed that ROR2(+) cells produce mesoderm progeny, APA(+) cells generate syncytiotrophoblasts and CD87(+) cells give rise to vasculature. The same progenitor classes emerged during the differentiation of human induced pluripotent stem cells (hiPSCs). These markers and progenitors provide tools for purifying human tissue-regenerating progenitors and for studying the commitment of pluripotent stem cells to lineage progenitors.
为了鉴定源自人类胚胎干细胞（hESC）的早期定向祖细胞，我们用 >400 种识别细胞表面抗原的抗体对自我更新、BMP4 处理和维甲酸处理的培养物进行了筛选。对 >30 个亚群进行排序，然后对发育基因进行转录分析，确定了四个不同的候选祖细胞群。在自我更新的培养物中检测到的亚群，包括 CXCR4(+)细胞，表达原始内胚层基因。在小鼠胚胎的内脏内胚层中证实了 Cxcr4 在原始内胚层中的表达。BMP4 诱导的祖细胞表现出中胚层、滋养层和血管内皮细胞的基因特征，分别提示与原肠胚阶段原始条纹、绒毛膜和尿囊前体相对应。体外和体内的功能研究证实，ROR2(+)细胞产生中胚层祖细胞，APA(+)细胞产生合体滋养层细胞，CD87(+)细胞产生血管。在人类诱导多能干细胞（hiPSC）的分化过程中也出现了相同的祖细胞群。这些标志物和祖细胞为纯化人类组织再生祖细胞以及研究多能干细胞向谱系祖细胞的定向提供了工具。
