Lo Nigro Antonio, de Jaime-Soguero Anchel, Khoueiry Rita, Cho Dong Seong, Ferlazzo Giorgia Maria, Perini Ilaria, Abon Escalona Vanesa, Aranguren Xabier Lopez, Chuva de Sousa Lopes Susana M, Koh Kian Peng, Conaldi Pier Giulio, Hu Wei-Shou, Zwijsen An, Lluis Frederic, Verfaillie Catherine M
Department of Development and Regeneration, Stem Cell Biology and Embryology, KU Leuven Stem Cell Institute, Herestraat 49, Onderwijs en Navorsing 4, Box 804, 3000 Leuven, Belgium; Ri.Med Foundation, Department of Laboratory Medicine and Advanced Biotechnologies, IRCCS-ISMETT (Istituto Mediterraneo per i Trapianti e Terapie ad Alta Specializzazione), Via Tricomi 5, 90127 Palermo, Italy.
Department of Development and Regeneration, Stem Cell Biology and Embryology, KU Leuven Stem Cell Institute, Herestraat 49, Onderwijs en Navorsing 4, Box 804, 3000 Leuven, Belgium.
Stem Cell Reports. 2017 Feb 14;8(2):318-333. doi: 10.1016/j.stemcr.2016.12.010. Epub 2017 Jan 12.
In early mouse pre-implantation development, primitive endoderm (PrE) precursors are platelet-derived growth factor receptor alpha (PDGFRα) positive. Here, we demonstrated that cultured mouse embryonic stem cells (mESCs) express PDGFRα heterogeneously, fluctuating between a PDGFRα+ (PrE-primed) and a platelet endothelial cell adhesion molecule 1 (PECAM1)-positive state (epiblast-primed). The two surface markers can be co-detected on a third subpopulation, expressing epiblast and PrE determinants (double-positive). In vitro, these subpopulations differ in their self-renewal and differentiation capability, transcriptional and epigenetic states. In vivo, double-positive cells contributed to epiblast and PrE, while PrE-primed cells exclusively contributed to PrE derivatives. The transcriptome of PDGFRα subpopulations differs from previously described subpopulations and shows similarities with early/mid blastocyst cells. The heterogeneity did not depend on PDGFRα but on leukemia inhibitory factor and fibroblast growth factor signaling and DNA methylation. Thus, PDGFRα cells represent the in vitro counterpart of in vivo PrE precursors, and their selection from cultured mESCs yields pure PrE precursors.
在小鼠植入前早期发育过程中，原始内胚层（PrE）前体呈血小板衍生生长因子受体α（PDGFRα）阳性。在此，我们证明培养的小鼠胚胎干细胞（mESCs）异质性表达PDGFRα，在PDGFRα阳性（PrE启动）状态和血小板内皮细胞黏附分子1（PECAM1）阳性状态（上胚层启动）之间波动。这两种表面标志物可在表达上胚层和PrE决定簇的第三个亚群（双阳性）上共同检测到。在体外，这些亚群在自我更新和分化能力、转录和表观遗传状态方面存在差异。在体内，双阳性细胞对形成上胚层和PrE有贡献，而PrE启动细胞仅对PrE衍生物有贡献。PDGFRα亚群的转录组不同于先前描述的亚群，与早期/中期囊胚细胞有相似之处。这种异质性不依赖于PDGFRα，而是依赖于白血病抑制因子和成纤维细胞生长因子信号传导以及DNA甲基化。因此，PDGFRα细胞代表体内PrE前体的体外对应物，从培养的mESCs中筛选它们可产生纯PrE前体。
