Pig Epiblast Stem Cells Depend on Activin/Nodal Signaling for Pluripotency and Self-Renewal
Pig Epiblast Stem Cells Depend on Activin/Nodal Signaling for Pluripotency and Self-Renewal
复制标题
DOI：
10.1089/scd.2010.0012
复制
发表时间：
2010-10-01
影响因子：
4
通讯作者：
Allegrucci, Cinzia
中科院分区：
文献类型：
--
作者：
Alberio, Ramiro;Croxall, Nicola;Allegrucci, Cinzia
激活素/Nodal 信号传导是维持小鼠外胚层干细胞和人胚胎干细胞 (hESC) 的多能性和自我更新所必需的。在这项研究中，我们研究了这种信号传导机制是否也适用于源自 10.5-12 天猪胚胎的培养外胚层。在小鼠饲养层和补充有碱性成纤维细胞生长因子（bFGF）的培养基上建立猪外胚层干细胞系（pEpiSC）。 pEpiSC 表达核心多能因子 OCT4（或 POU5F1）、NANOG、SOX2 和 NODAL，但不表达 REX1 或碱性磷酸酶活性。通过添加特异性 JAK I 抑制剂 420099 和抗 LIF 抗体超过 3 代来阻断白血病抑制因子 (LIF)/JAK/STAT3 通路不会影响 pEpiSC 的多能性。相比之下，使用 Alk-5 抑制剂 SB431542（阻断 Activin/Nodal 通路）生长的细胞很容易向神经谱系分化。根据其向外胚层、中胚层和内胚层的分化潜力，pEpiSC 具有多能性。响应骨形态发生蛋白 4 (BMP-4)，这些细胞可被诱导分化为滋养外胚层和生殖细胞前体。总之，我们的研究表明猪外胚层表达核心多能性基因，并且 pEpiSC 维持自我更新的能力取决于 Activin/Nodal 信号传导。这项研究提供了进一步的证据，证明通过 Activin/Nodal 信号维持多能性在哺乳动物中是保守的。
Activin/Nodal signaling is required for maintaining pluripotency and self-renewal of mouse epiblast stem cells and human embryonic stem cells (hESC). In this study, we investigated whether this signaling mechanism is also operative in cultured epiblasts derived from Days 10.5-12 pig embryos. Pig epiblast stem cell lines (pEpiSC) were established on mouse feeder layers and medium supplemented with basic fibroblast growth factor (bFGF). pEpiSC express the core pluripotency factors OCT4 (or POU5F1), NANOG, SOX2, and NODAL, but they do not express REX1 or alkaline phosphatase activity. Blocking leukemia inhibitory factor (LIF)/JAK/STAT3 pathway by adding the specific JAK I inhibitor 420099 and an anti-LIF antibody over 3 passages did not affect pluripotency of pEpiSC. In contrast, cells grown with the Alk-5 inhibitor SB431542, which blocks Activin/Nodal pathway, differentiated readily toward the neural lineage. pEpiSC are pluripotent, as established by their differentiation potential to ectoderm, mesoderm, and endoderm. These cells can be induced to differentiate toward trophectoderm and to germ cell precursors in response to bone morphogenetic protein 4 (BMP-4). In conclusion, our study demonstrates that pig epiblasts express the core pluripotency genes and that the capacity for maintaining self-renewal in pEpiSC depends on Activin/Nodal signaling. This study provides further evidence that maintenance of pluripotency via Activin/Nodal signal is conserved in mammals.