Characterization and multilineage differentiation of embryonic stem cells derived from a buffalo parthenogenetic embryo

Hathaitip Sritanaudomchai, Kanok Pavasuthipaisit, Yindee Kitiyanant, Piengchai Kupradinun, Shoukhrat Mitalipov, Thanit Kusamran

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    Abstract

    Embryonic stem (ES) cells derived from mammalian embryos have the ability to form any terminally differentiated cell of the body. We herein describe production of parthenogenetic buffalo (Bubalus Bubalis) blastocysts and subsequent isolation of an ES cell line. Established parthenogenetic ES (PGES) cells exhibited diploid karyotype and high telomerase activity. PGES cells showed remarkable long-term proliferative capacity providing the possibility for unlimited expansion in culture. Furthermore, these cells expressed key ES cell-specific markers defined for primate species including stage-specific embryonic antigen-4 (SSEA-4), tumor rejection antigen-1-81 (TRA-1-81), and octamer-binding transcription factor 4 (Oct-4). In vitro, in the absence of a feeder layer, cells readily formed embryoid bodies (EBs). When cultured for an extended period of time, EBs spontaneously differentiated into derivatives of three embryonic germ layers as detected by PCR for ectodermal (nestin, oligodendrocytes, and tubulin), mesodermal (scleraxis, α-skeletal actin, collagen II, and osteocalcin) and endodermal markers (insulin and α-fetoprotein). Differentiation of PGES cells toward chondrocyte lineage was directed by supplementing serum-containing media with ascorbic acid, β-glycerophosphate, and dexamethasone. Moreover, when PGES cells were injected into nude mice, teratomas with derivatives representing all three embryonic germ layers were produced. Our results suggest that the cell line isolated from a parthenogenetic blastocyst holds properties of ES cells, and can be used as an in vitro model to study the effects of imprinting on cell differentiation and as an a invaluable material for extensive molecular studies on imprinted genes.

    Original languageEnglish (US)
    Pages (from-to)1295-1302
    Number of pages8
    JournalMolecular Reproduction and Development
    Volume74
    Issue number10
    DOIs
    StatePublished - Oct 1 2007

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    Keywords

    • Buffalo
    • Embryonic stem cells
    • Parthenogenesis
    • Pluripotent

    ASJC Scopus subject areas

    • Genetics
    • Developmental Biology
    • Cell Biology

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