title: Embryos aggregation improves development and imprinting gene expression in mouse parthenogenesis.
abstract: Mouse parthenogenetic embryonic stem cells (PgESCs) 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 a2 PgESCs) had a higher efficiency than that of PgESCs, 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 PgESCs. 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-a4 PgESCs 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 (fESCs). In addition, we acquired a 13.5-day fetus totally derived from GFP-a4 PgESCs 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 PgESCs. It implied that a4 PgESCs could serve as a better scientific model applied in translational medicine and imprinting gene study.
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, China.
authors.forename: Guang-Yu
authors.initials: GY
authors.lastname: Bai
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, China.
authors.forename: Si-Hang
authors.initials: SH
authors.lastname: Song
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, China.
authors.forename: Zhen-Dong
authors.initials: ZD
authors.lastname: Wang
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, China.
authors.forename: Zhi-Yan
authors.initials: ZY
authors.lastname: Shan
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, China.
authors.forename: Rui-Zhen
authors.initials: RZ
authors.lastname: Sun
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, China.
authors.forename: Chun-Jia
authors.initials: CJ
authors.lastname: Liu
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, China.
authors.forename: Yan-Shuang
authors.initials: YS
authors.lastname: Wu
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, China.
authors.forename: Tong
authors.initials: T
authors.lastname: Li
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Lei
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D001757:Blastomeres / Q000166:cytology / Q000378:metabolism*; D002449:Cell Aggregation / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics; D047108:Embryonic Development / Q000235:genetics; D005260:Female; D005455:Fluorescent Antibody Technique; D018507:Gene Expression Regulation, Developmental; D018392:Genomic Imprinting / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D010312:Parthenogenesis / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors
keywords: aggregation; cells; imprinting gene; mouse parthenogenetic embryonic stem; mouse parthenogenetic embryos; post-implantation development