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Parthenogenetically derived mammalian embryos, with no paternal genome, are not viable and die, largely from defective placental growth attributed to a lack of paternal effect, resulting in the low blastulation rate and low derivation efficiency of parthenogenetic embryonic stem cells (pESCs). | Turkish Journal of Biology http://journals.tubitak.gov.tr/biology/ Research Article Turk J Biol (2017) 41: 629-639 © TÜBİTAK doi:10.3906/biy-1612-30 Aggregation of a parthenogenetic diploid embryo and a male embryo improves the blastocyst development and parthenogenetic embryonic stem cell derivation Xiaoyan QIU*, Nan LI*, Xiong XIAO, Yuemin LI** Embryo Engineering Lab, College of Animal Science & Technology, Southwest University, Chong Qing, P.R. China Received: 14.12.2016 Accepted/Published Online: 11.04.2017 Final Version: 24.08.2017 Abstract: Parthenogenetically derived mammalian embryos, with no paternal genome, are not viable and die, largely from defective placental growth attributed to a lack of paternal effect, resulting in the low blastulation rate and low derivation efficiency of parthenogenetic embryonic stem cells (pESCs). Therefore, the present study, by the optimization of parthenogenetic embryo production and the aggregation of the parthenogenetic diploid embryo and the identified male embryo, aims to investigate a method to improve the development of parthenogenetic embryo and pESC derivation in mice. Using different chemical combinations for the optimization, we found that the heterozygous diploid type had a significantly higher blastulation rate than the haploid type (P 0.05; Table 3). Oocytes activated by SrCl2 combined with CB for 4 h produced the highest amount of the diploid type (70.4%; Table 3) and the highest blastulation rate (29.6%; Table 3). The in vitro development of parthenogenetic embryos is seen in Figure 4. 3.2. Sex identification for male embryos 3.2.1. Optimization of duplex PCR method for sex identification The duplex PCR amplification results (Figure 5) showed that there were two bands in Lanes 1, 2, 3, and 7 (4 male mice): one band of 250 bp in length and another band between 250 bp and 500 bp, which were consistent with the expected lengths of 250 bp and 399 bp. There was only one band in Lanes 4, 5, 6, and 8 (4 female
