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Establishment of mouse androgenetic embryonic stem cells by double sperm injection and differentiation into beating embryoid body

Published online by Cambridge University Press:  23 September 2019

Lei Lei Open the ORCID record for Lei Lei [Opens in a new window] ,
Lili Hu ,
Tong Li ,
Xinghui Shen ,
Xiao Liang ,
Yajun Chen ,
Xiuqing Feng ,
Zhiwen Yang  and
Zhiyan Shan
Lei Lei*
Affiliation:
Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, Heilongjiang, China Key Laboratory of Preservation of Human Genetic Resources and Disease Control in China (Harbin Medical University), Ministry of Education, China
Lili Hu
Affiliation:
Department of Reproductive Medicine, the First People’s Hospital of Jining City, Jining, 272100, Shandong, China
Tong Li
Affiliation:
Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, Heilongjiang, China
Xinghui Shen
Affiliation:
Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, Heilongjiang, China
Xiao Liang
Affiliation:
Department of Cardiology, the First Affiliated Hospital of Harbin Medical University, Harbin, 150081, Heilongjiang, China
Yajun Chen
Affiliation:
Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, Heilongjiang, China
Xiuqing Feng
Affiliation:
Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, Heilongjiang, China
Zhiwen Yang
Affiliation:
Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, Heilongjiang, China
Zhiyan Shan
Affiliation:
Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, Heilongjiang, China
*
Address for correspondence: Lei Lei. Department of Histology and Embryology, Harbin Medical University, 194 Xuefu Road, Nangang District, Harbin 150081, Heilongjiang, China, Tel: +86 451 86674518. Fax: +86 451 87503325. E-mail: lei086@ems.hrbmu.edu.cn
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Summary

Androgenetic embryonic stem (AgES) cells offer a possible tool for patient-specific pluripotent stem cells that will benefit genomic imprinting studies and clinic applications. However, the difficulty in producing androgenetic embryos and the unbalanced expression of imprinted genes make the therapeutic applicability of AgES cells uncertain. In this study, we produced androgenetic embryos by injecting two sperm into an enucleated metaphase II (MII) oocyte. By this method, 88.48% of oocytes survived after injection, and 20.24% of these developed to the blastocyst stage. We successfully generated AgES cell lines from the androgenetic embryos and assayed the expression of imprinted genes in the cell lines. We found that the morphological characteristics of AgES cells were similar to that of fertilized embryonic stem cells (fES), such as expression of key pluripotent markers, and generation of cell derivatives representing all three germ layers following in vivo and in vitro differentiation. Furthermore, activation of paternal imprinted genes was detected, H19, ASC12 and Tss3 in AgES cell activation levels were lower while other examined genes showed no significant difference to that of fES cells. Interestingly, among examined maternal imprinted genes, only Mest and Igf2 were significantly increased, while levels of other detected genes were no different to that of fES cells. These results demonstrated that activation of some paternal imprinted genes, as well as recovery of maternal imprinted genes, was present in AgES cells. We differentiated AgES cells into a beating embryoid body in vitro, and discovered that the AgES cells did not show significant higher efficiency in myocardial differentiation potential.

Keywords

Androgenetic embryonic stem cellsBeating embryoid bodies (EB)Double sperms injectionImprinted genesMouse
Type
Research Article
Information
Zygote , Volume 27 , Issue 6 , December 2019 , pp. 405 - 412
Copyright
© Cambridge University Press 2019 

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