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| Inter-species Cloned Embryo Development with the Transcription Factor Induced Tibetan Antelope (Pantholops hodgsoni) Fibro Blast Cells |
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Abstract The Tibetan antelope (Pantholops hodgsoni) is endemic to the Tibetan Plateau and becomes endang ered species, which is national class Ⅰprotected animals in China. Inter-species somatic cell nuclear transfer (iSCNT) would be a possible rescue strategy for these species. iSCNT has been regarded as a potential alternative for rescuing highly endangered species and can be used as a model for studying nuclear-cytoplasmic interactions. However, iSCNT embryos often fail to produce viable offspring. In this study, the Tibetan antelope fibroblasts were treated by the 4 classic mouse (Mus musculus) transcription factors of octamer-binding transcription factor 4 (OCT-4), sex determining region Y -Box 2 (SOX2), Kruppel-like factor 4(KLF4) and (avian myelocytomatosis viral oncogene homolog (C-MYC) to induce the cell reprogramming. The pluripotent gene expression, cell growth curve and cell cycle of the induced cells were detected. The induced cells (iPS-ZLY) were then used as donors to transfer to the enucleated bovine (Bos taurus) oocytes. The results showed that the induced Tibetan antelope fibroblast cells (iPS-ZLY) had numerically higher cell mass in the G2-M stage than the Tibetan antelope fibroblasts (ZLY)(21.5% vs 16.7%). Cell growth curve indicated that iPS-ZLY cells proliferated faster than the ZLY cells, iPS - ZLY cells gone into the growth of the plateau in 4 d, while the ZLY cells gone into a plateau in 6 d. The karyotypes of both iPS-ZLY and ZLY cells were similar (68.3%vs 69.6%). The pluripotent gene OCT-4 expressed only in the iPS-ZLY. When transfered of the iPS-ZLY and ZLY cells, irrespectively to enucleated bovine oocytes, the iPS-ZLY cells resulted in 2.4% cloned blastocyst development, while the ZLY cells yielded 0.95% cloned blasatocysts. These results suggested that treatment of Tibetan antelope fibroblast cells with transcriptional factors enhanced cell proliferation, induced pluripotent gene expression and improved inter-species cloned embryo development. This was the first report using the 4 mouse transcription factors to induce the Tibetan antelope somatic cells, and then to evaluate the potential development of the iSCNT reconstructed embryos, resulted from the induced cells. This research provides an alternative method to increase the reprogramming of endangered animal cells and probably is beneficial for endangered animal protection.
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Received: 05 January 2016
Published: 20 May 2016
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