Embryos aggregation improves development and imprinting gene expression in mouse parthenogenesis
Autor: | Chun-Jia Liu, Ruizhen Sun, Zhiyan Shan, Zhen-Dong Wang, Guang-Yu Bai, Lei Lei, Si-Hang Song, Yanshuang Wu, Tong Li |
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Rok vydání: | 2016 |
Předmět: |
0301 basic medicine
Blastomeres Time Factors Cellular differentiation Parthenogenesis Embryonic Development Fluorescent Antibody Technique Mice Transgenic Biology Genomic Imprinting Mice 03 medical and health sciences medicine Animals Blastocyst Zona pellucida Cell Aggregation Genetics Reverse Transcriptase Polymerase Chain Reaction Gene Expression Regulation Developmental Cell Differentiation Mouse Embryonic Stem Cells Embryo Cell Biology Embryonic stem cell Cell aggregation Cell biology 030104 developmental biology medicine.anatomical_structure embryonic structures Female Stem cell Genomic imprinting Developmental Biology |
Zdroj: | Development, Growth & Differentiation. 58:270-279 |
ISSN: | 0012-1592 |
Popis: | 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. |
Databáze: | OpenAIRE |
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