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首页> 外文期刊>Cell cycle >The crucial role of Activin A on the formation of primordial germ cell-like cells from skin-derived stem cells in vitro
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The crucial role of Activin A on the formation of primordial germ cell-like cells from skin-derived stem cells in vitro

机译：激活素A在体外从皮肤干细胞形成原始生殖细胞样细胞中的关键作用

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Primordial germ cells (PGCs) are founder cells of the germ cell lineage, and can be differentiated from stem cells in an induced system in vitro. However, the induction conditions need to be optimized in order to improve the differentiation efficiency. Activin A (ActA) is a member of the TGF- super family and plays an important role in oogenesis and folliculogenesis. In the present study, we found that ActA promoted PGC-like cells (PGCLCs) formation from mouse skin-derived stem cells (SDSCs) in both embryoid body-like structure (EBLS) differentiation and the co-culture stage in a dose dependent manner. ActA treatment (100ng/ml) during EBLS differentiation stage and further co-cultured for 6days without ActA significantly increased PGCLCs from 53.2% to 82.8%, and as well as EBLS differentiation without ActA followed by co-cultured with 100ng/ml ActA for 4 to 12days with the percentage of PGCLCs increasing markedly in vitro. Moreover, mice treated with ActA at 100ng/kg body weight from embryonic day (E) 5.5-12.5 led to more PGCs formation. However, the stimulating effects of ActA were interrupted by Smad3 RNAi, and in an in vitro cultured Smad3(-/-) mouse skin cells scenario. SMAD3 is thus likely a key effecter molecule in the ActA signaling pathway. In addition, we found that the expression of some epiblast cell markers, Fgf5, Dnmt3a, Dnmt3b and Wnt3, was increased in EBLSs cultured for 4days or PGCLCs co-cultured for 12days with ActA treatment. Interestingly, at 16days of differentiation, the percentage of PGCLCs was decreased in the presence of ActA, but the expression of meiosis-relative genes, such as Stra8, Dmc1, Sycp3 and Sycp1, was increased. In conclusion, our data here demonstrated that ActA can promote PGCLC formation from SDSCs in vitro, at early stages of differentiation, and affect meiotic initiation of PGCLCs in later stages
机译：原始生殖细胞（PGC）是生殖细胞谱系的基础细胞，可以在体外诱导系统中与干细胞区分开。但是，需要优化诱导条件以提高分化效率。激活素A（ActA）是TGF-super家族的成员，在卵子发生和卵泡发生中起重要作用。在本研究中，我们发现ActA以类鼠体样结构（EBLS）分化和共培养阶段促进了小鼠皮肤衍生干细胞（SDSCs）形成的PGC样细胞（PGCLC）的形成，并呈剂量依赖性。 。在EBLS分化阶段进行ActA处理（100ng / ml），并在不使用ActA的情况下进一步共培养6天，PGCLCs将从53.2％增至82.8％，并且在不使用ActA的情况下将EBLS分化程度提高，然后与100ng / ml的ActA共培养4次。到12天，PGCLC的百分比在体外显着增加。此外，从胚胎日（E）5.5-12.5开始，以Acta以100ng / kg体重处理的小鼠导致更多的PGC形成。但是，ActA的刺激作用被Smad3 RNAi中断，并在体外培养的Smad3（-/-）小鼠皮肤细胞中发生。因此，SMAD3可能是ActA信号通路中的关键效应分子。此外，我们发现在培养4天的EBLS或通过ActA处理共培养12天的PGCLCs中，某些表皮细胞标记Fgf5，Dnmt3a，Dnmt3b和Wnt3的表达增加。有趣的是，在分化的第16天，ActA存在时PGCLC的百分比降低，但减数分裂相关基因如Stra8，Dmc1，Sycp3和Sycp1的表达增加。总之，我们的数据表明ActA可以在分化的早期促进体外SDSCs形成PGCLC，并在后期影响PGCLC的减数分裂启动

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