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Embryos aggregation improves development and imprinting gene expression in mouse parthenogenesis
Author(s) -
Bai GuangYu,
Song SiHang,
Wang ZhenDong,
Shan ZhiYan,
Sun RuiZhen,
Liu ChunJia,
Wu YanShuang,
Li Tong,
Lei Lei
Publication year - 2016
Publication title -
development, growth and differentiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 66
eISSN - 1440-169X
pISSN - 0012-1592
DOI - 10.1111/dgd.12271
Subject(s) - genomic imprinting , blastocyst , imprinting (psychology) , biology , parthenogenesis , embryo , embryonic stem cell , embryogenesis , gene , andrology , genetics , microbiology and biotechnology , stem cell , gene expression , dna methylation , medicine
Mouse parthenogenetic embryonic stem cells (Pg ESC s) could be applied to study imprinting genes and are used in cell therapy. Our previous study found that stem cells established by aggregation of two parthenogenetic embryos at 8‐cell stage (named as a 2 Pg ESC s) had a higher efficiency than that of Pg ESC s, and the paternal expressed imprinting genes were observably upregulated. Therefore, we propose that increasing the number of parthenogenetic embryos in aggregation may improve the development of parthenogenetic mouse and imprinting gene expression of Pg ESC s. To verify this hypothesis, we aggregated four embryos together at the 4‐cell stage and cultured to the blastocyst stage (named as 4aPgB). qPCR detection showed that the expression of imprinting genes Igf2, Mest, Snrpn, Igf2r, H19, Gtl2 in 4aPgB were more similar to that of fertilized blastocyst (named as fB ) compared to 2aPgB (derived from two 4‐cell stage parthenogenetic embryos aggregation) or PgB (single parthenogenetic blastocyst). Post‐implantation development of 4aPgB extended to 11 days of gestation. The establishment efficiency of GFP ‐a 4 Pg ESC s which derived from GFP ‐4aPgB is 62.5%. Moreover, expression of imprinting genes Igf2, Mest, Snrpn, notably downregulated and approached the level of that in fertilized embryonic stem cells ( fESC s). In addition, we acquired a 13.5‐day fetus totally derived from GFP ‐a 4 Pg ESC s with germline contribution by 8‐cell under zona pellucida ( ZP ) injection. In conclusion, four embryos aggregation improves parthenogenetic development, and compensates imprinting genes expression in Pg ESC s. It implied that a 4 Pg ESC s could serve as a better scientific model applied in translational medicine and imprinting gene study.