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“Conservation cloning” of vulnerable Esfahan mouflon (Ovis orientalis isphahanica): in vitro and in vivo studies

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Abstract

Among the wide range of bio-conservational strategies envisaged, recent accomplishments in the field of interspecies somatic cell nuclear transfer (iSCNT) hold considerable promise due to its unique potential to decelerate or prevent rapid loss of animal genetic resources, and even to revive extinct species. Accordingly, this study was carried out to investigate if in vitro matured and enucleated oocytes of domestic sheep could be used for interspecies conservation cloning of Esfahan mouflon (Ovis orientalis isphahanica), a vulnerable species classified by the International Union for Conservation of Nature. Cryo-banked fibroblasts of a mouflon (derived from a genome resource bank) and a domestic sheep (prepared during a recent study) were cultured in vitro and used for karyotyping. Prior to SCNT, fibroblast donor cells were serum starved for 5 days. Using the zona-free SCNT technique, in vitro matured and enucleated domestic sheep oocytes were reconstituted with nuclei donor cells of mouflon and domestic sheep, and their competencies for in vitro development to the blastocyst stage were compared. The cloned mouflon blastocysts were then surgically transferred into the uterus of the synchronized domestic sheep. Karyotype analysis confirmed that fibroblasts of the Esfahan mouflon had the correct number of diploid chromosomes (2n = 54). Evaluation of 907 activated reconstructs [Esfahan mouflon (n = 667), domestic sheep (n = 240)] revealed no significant difference in the term of blastocyst development (7.6 ± 0.5% vs. 9.3 ± 0.5%, respectively). After the transfer of 12 cloned Esfahan mouflon blastocysts to five domestic sheep recipients, two (40.0%) pregnancies were established in which both (100%) were sustained until caesarean section (days 147 and 150 of pregnancy, respectively) and culminated in the live births of cloned Esfahan mouflon lambs. However, the newborns did not survive and died soon after birth. Karyotype and genetic analyses confirmed that both clones had correct diploid chromosome number (2n = 54), and were genetically identical to each other in addition to their original cell donor. This study highlighted the importance of “conservation cloning” using closely related abundant alternate species.

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Acknowledgments

We would like to thank the Department of Environment of the Islamic Republic of Iran, Isfahan Province, for their full support as well as providing the opportunity and samples for carrying out this study. The authors also would like to thank Mrs. Hajheidari for her help with neonatal care. This study was funded by a grant from the Royan Institute of IRI.

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Authors and Affiliations

  1. Department of Reproduction and Development, Royan Institute for Animal Biotechnology, ACECR, Isfahan, Iran

    M. Hajian, S. M. Hosseini, P. Abedi, S. Ostadhosseini, L. Hosseini, F. Moulavi & M. H. Nasr-Esfahani

  2. Department of Animal Science, Marvdasht Branch, Islamic Azad University, Marvdasht, Iran

    M. Forouzanfar

  3. Department of Genetics, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

    H. Gourabi

  4. Department of Reproduction and Development, Cell Sciences Research Centre, ACECR, Isfahan, Iran

    A. H. Shahverdi & A. Amiri-Yekta

  5. Department of Infertility Imaging, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

    A. Vosough Taghi Dizaj

  6. Department of Andrology, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

    A. Vosough Taghi Dizaj

  7. Isfahan Fertility and Infertility Centre, Isfahan, Iran

    S. A. Kalantari & Z. Fotouhi

  8. Department of Medical Sciences, Isfahan University, Isfahan, Iran

    R. Iranpour

  9. Imam Mousaa-e-Kazem Burn Hospital, Isfahan, Iran

    H. Mahyar

  10. Department of Reproduction and Development, Cell Sciences Research Centre, Royan Institute for Animal Biotechnology, ACECR, Isfahan, Iran

    M. H. Nasr-Esfahani

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  1. M. Hajian
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  2. S. M. Hosseini
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  3. M. Forouzanfar
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  4. P. Abedi
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  5. S. Ostadhosseini
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  6. L. Hosseini
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  7. F. Moulavi
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  9. A. H. Shahverdi
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  16. M. H. Nasr-Esfahani
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Correspondence to M. H. Nasr-Esfahani.

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Communicated by C. Gortázar

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Hajian, M., Hosseini, S.M., Forouzanfar, M. et al. “Conservation cloning” of vulnerable Esfahan mouflon (Ovis orientalis isphahanica): in vitro and in vivo studies. Eur J Wildl Res 57, 959–969 (2011). https://doi.org/10.1007/s10344-011-0510-5

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  • DOI: https://doi.org/10.1007/s10344-011-0510-5

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