To improve the isolation efficiency of parthenogenetic embryonic stem cells (pESCs) in mice, it is necessary to optimize the method to increase in vitro developmental competence of mice parthenogenetic blastocysts. Therefore, this study aims to investigate an optimal method for the production of mouse parthenogenetic blastocysts and isolation of pESC colonies by comparing the effects of two methods: (1) the treatment of histone deacetylase inhibitor PXD101 before, during, or after parthenogenetic activation; (2) parthenogenetic embryo aggregation; and (3) their combination treatment. The results suggest that application of PXD101 treatment and embryo aggregation could both improve the development of mouse parthenogenetic blastocysts (50 nM PXD101 treated 4 hours during activation and further 4 hours after activation: 40.0% vs. 20.0%; p < 0.05; two-cell embryo aggregation: 38.3% vs. 20.0%; p < 0.05) and also enhance the isolation rate of pESC colonies (PXD101: 33.3% vs. 11.8%; p < 0.05; two-cell embryo aggregation: 36.4% vs. 11.8%; p < 0.05). The combination of their treatments had the higher rate of parthenogenetic blastocyst development (41.7%) and significantly higher rate of pESC colony isolation from parthenogenetic blastocysts (45.0%); therefore, we concluded that the combination of these two methods (50 nM PXD101 treated for 8 hours and then aggregated at two-cell stage with 0.25% pronase for 10 minutes in our self-made concave) is considered the optimal way for the in vitro development of parthenogenetic blastocysts and subsequent pESC colony isolation in mice, opening new opportunities for application of this combination method to improve the parthenogenetic embryo development in other species.

中文翻译：

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PXD101和胚胎聚集对小鼠孤雌生殖胚胎体外发育的影响。

为了提高小鼠孤雌生殖胚胎干细胞（pESCs）的分离效率，有必要优化方法以提高小鼠孤雌生殖胚泡的体外发育能力。因此，本研究旨在通过比较两种方法的效果，研究产生小鼠孤雌性胚泡和分离pESC菌落的最佳方法：（1）组蛋白脱乙酰基酶抑制剂PXD101在孤雌性激活之前，之中或之后的治疗；（2）孤雌胚胎聚集；（三）联合治疗。结果表明，应用PXD101处理和胚胎聚集均可改善小鼠孤雌生殖胚泡的发育（激活期间4小时和激活后4小时分别处理50 nM PXD101：40.0％vs. 20.0％； p <0.05； P <0.05）。两细胞胚胎聚集：38.3％和20.0％；p <0.05）并提高了pESC集落的分离率（PXD101：33.3％vs. 11.8％; p <0.05;两细胞胚胎聚集：36.4％vs. 11.8％; p <0.05）。他们的联合治疗具有更高的孤雌生殖胚泡发育率（41.7％）和显着更高的从孤雌生殖胚囊中分离出pESC菌落的率（45.0％）；因此，我们得出结论，这两种方法的组合（将50 nM PXD101处理8小时，然后在两细胞阶段以0.25％的链霉蛋白酶在我们自制的凹面中聚集10分钟）被认为是体外的最佳方法小鼠孤雌生殖胚泡的发育和随后的pESC集落分离，