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| 1 | 激活素A通过TGFβ/SMAD信号通路促进鸡原始生殖细胞增殖显示文摘原始生殖细胞(primordial germ cells,PGCs)是一类具有分化潜能的生殖细胞。为了检测激活素A(activin A)对鸡胚血液PGCs体外存活和增殖的影响,本研究从第13~15期鸡胚血液中分离提取PGCs,并进行鉴定;在完全培养液中分别添加0和100 ng/m L activin A培养48 h后,检测PGCs数目、形态及细胞周期;检测Dazl(deleted in azoospermia-like)基因甲基化水平;并通过Western blot检测转化生长因子-β(transforming growth factor-β,TGF-β)信号通路中TGF-β、SMAD2/3的表达和磷酸化变化。结果显示,从鸡胚血液中分离的PGCs,经过红细胞裂解液处理和传代后,可获得高纯度PGCs,且具有良好的生长趋势。培养的PGCs呈阶段特异性胚胎抗原(stage specific embryonic antigen 1,SSEA-1)、鸡(Gallus gallus)VASA同源基因(chicken vasa homologue,CVH)免疫染色和过碘酸希夫(periodic acid schiff reaction,PAS)反应染色阳性,并且表达多能性相关基因Pou V、Nanog和Sox2(SRY(sex determining region Y)-box 2),表明经过体外培养后,鸡胚血液PGCs仍能维持其生物学特性。添加activin A,可显著降低PGCs中Dazl基因甲基化水平,提高PGCs的体外存活能力。activin A处理后,显著增加了PGCs的数量;上调细胞周期蛋白(cyclins)及细胞周期蛋白依赖性激酶(cyclin-dependent kinases,CDKs)的表达;流式细胞检测进一步证实了activin A增加S/G2期PGCs细胞的比例。activin A显著上调了TGF-β蛋白的表达,促进SMAD2/3的磷酸化。以上结果表明,activin A可通过激活TGF-β/SMAD信号通路显著提高PGCs的体外存活和增殖能力。研究结果为揭示activin A对鸡胚PGCs增殖的作用机制提供了依据,为利用PGCs体外培养模型进一步深入研究生殖细胞发育调控提供基础资料。 | 于敏莉 李铭 虞德兵 杜文兴 | 2017 | 农业生物技术学报2017，25，4： | 2 |
| 2 | Formative pluripotent stem cells show features of epiblast cells poised for gastrulation显示文摘The pluripotency of mammalian early and late epiblast could be recapitulated by naïve embryonic stem cells(ESCs)and primed epiblast stem cells(EpiSCs),respectively.However,these two states of pluripotency may not be sufficient to reflect the full complexity and developmental potency of the epiblast during mammalian early development.Here we report the establishment of self-renewing formative pluripotent stem cells(fPSCs)which manifest features of epiblast cells poised for gastrulation.fPSCs can be established from different mouse ESCs,pre-/early-gastrula epiblasts and induced PSCs.Similar to pre-/early-gastrula epiblasts,fPSCs show the transcriptomic features of formative pluripotency,which are distinct from naïve ESCs and primed EpiSCs.fPSCs show the unique epigenetic states of E6.5 epiblast,including the super-bivalency of a large set of developmental genes.Just like epiblast cells immediately before gastrulation,fPSCs can efficiently differentiate into three germ layers and primordial germ cells(PGCs)in vitro.Thus,fPSCs highlight the feasibility of using PSCs to explore the development of mammalian epiblast. | Xiaoxiao Wang Yunlong Xiang Yang Yu Ran Wang Yu Zhang Qianhua Xu Hao Sun Zhen-Ao Zhao Xiangxiang Jiang Xiaoqing Wang Xukun Lu Dandan Qin Yujun Quan Jiaqi Zhang Ng Shyh-Chang Hongmei Wang Naihe Jing Wei Xie Lei Li | 2021 | Cell Research2021，31，5： | 2 |
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