Generation and characterization of mouse parthenogenetic embryonic stem cells containing genomes from non-growing and fully grown oocytes

Authors

  • Hua Shao,

    1. Key Laboratory for Mammal Reproduction Biology and Biotechnology of Education Ministry, Inner Mongolia University, Hohhot 010021, China
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    • These authors contributed equally to this work.

  • Zhuying Wei,

    1. Key Laboratory for Mammal Reproduction Biology and Biotechnology of Education Ministry, Inner Mongolia University, Hohhot 010021, China
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    • These authors contributed equally to this work.

  • Lingling Wang,

    1. Key Laboratory for Mammal Reproduction Biology and Biotechnology of Education Ministry, Inner Mongolia University, Hohhot 010021, China
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    • These authors contributed equally to this work.

  • Lihua Wen,

    1. Department of Cellular and Molecular Medicine, University of Ottawa, Canada
    2. Ottawa Health Research Institute, Civic Campus, 725 Parkdale Avenue, Ottawa, ON K1Y 4E9, Canada
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  • Biao Duan,

    1. Key Laboratory for Mammal Reproduction Biology and Biotechnology of Education Ministry, Inner Mongolia University, Hohhot 010021, China
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  • Lie Mang,

    1. Key Laboratory for Mammal Reproduction Biology and Biotechnology of Education Ministry, Inner Mongolia University, Hohhot 010021, China
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  • Shorgan Bou

    Corresponding author
    1. Key Laboratory for Mammal Reproduction Biology and Biotechnology of Education Ministry, Inner Mongolia University, Hohhot 010021, China
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Corresponding author. Research Center for Laboratory Animals, Inner Mongolia University, 235 Daxuexilu, Hohhot 010021, China. Tel./fax: +86 471 4995071. shorganbou@163.com

Abstract

It is known that oocytes can be activated without male contribution in vitro and develop to blastocysts which are used to isolate parthenogenetic embryonic stem cells. Unfortunately, differentiation capacity of the parthenogenetic embryonic stem cells was rather lower than fertilized embryos derived ES cells, which might be the result of the absence of male genome. It had been found that some maternally expressed genes were repressed and some paternally expressed genes were expressed in the non-growing oocytes. Therefore, maternal genome from non-growing oocytes can partially act as “sperm genome”. In the present study, parthenogenetic blastocysts containing genome from non-growing and fully grown oocytes (named as NF-pBlastocysts) were produced by germinal vesicle transfer, and three newly established parthenogenetic embryonic stem (named as NF-pES) cell lines were derived from the resulting parthenogenetic blastocysts. All three NF-pES cell lines were positive for ES cell markers, such as alkaline phosphatase (AKP), stage-specific embryonic antigen 1 (SSEA-1) and octamer-binding transcription factor (Oct-4). They have a normal chromosome karyotype (40) and can be maintained in an undifferentiated state for extended periods of time. When NF-pES cells were injected into severe combined immunodeficient mice, teratomas with all three embryonic germ layers were obtained. The in vitro differentiation potential of NF-pES cells was analyzed by embryonic bodies (EB) formation. The expression of germ layer markers, such as nestin (ectoderm), desmin (mesoderm), and α-fetoprotein (endoderm) demonstrated that the NF-pES cells can differentiate into all three germ layers.

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