Journal of Animal Science and Technology

Korean Society of Animal Sciences and Technology (한국축산학회)
권호 표지
DOI QR코드

DOI QR Code

Production of transgenic cattle by somatic cell nuclear transfer (SCNT) with the human granulocyte colony-stimulation factor (hG-CSF)

  • Carvalho, Bruno P. (Institute of Biology, Universidade de Brasilia) ;
  • Cunha, Andrielle T.M. (Institute of Biology, Universidade de Brasilia) ;
  • Silva, Bianca D.M. (Laboratory of Animal Reproduction, Embrapa Genetic Resources and Biotechnology) ;
  • Sousa, Regivaldo V. (Laboratory of Animal Reproduction, Embrapa Genetic Resources and Biotechnology) ;
  • Leme, Ligiane O. (Department of Veterinary Medicine, Universidade Federal do Espirito Santo) ;
  • Dode, Margot A.N. (Institute of Biology, Universidade de Brasilia) ;
  • Melo, Eduardo O. (Laboratory of Animal Reproduction, Embrapa Genetic Resources and Biotechnology)
  • Received : 2018.12.04
  • Accepted : 2019.03.06
  • Published : 2019.03.31
Download PDF
⟨ Previous Next ⟩

Abstract

The hG-CSF (human Granulocyte Colony-Stimulating Factor) is a growth and stimulation factor capable of inducing the proliferation of bone marrow cells, several types of leukocytes, among other hematopoietic tissue cells. hG-CSF is used in used to treat anomalies that reder a small number of circulating white blood cells, which may compromise the immune defenses of the affected person. For these reasons, the production of hG-CSF in a bioreactor system using the mammary gland of genetic modified animals is a possibility of adding value to the bovine genetic material and reducing the costs of hG-CSF production in pharmaceutical industry. In this study, we aimed the production of transgenic hG-CSF bovine through the lipofection of bovine primary fibroblasts with an hG-CSF expression cassette and cloning these fibroblasts by the somatic cell nuclear transfer (SCNT) technique. The bovine fibroblasts transfected with the hG-CSF cassette presented a stable insertion of this construct into their genome and were efficiently synchronized to G0/G1 cell cycle stage. The transgenic fibroblasts were cloned by SCNT and produced 103 transferred embryos and 2 pregnancies, one of which reached 7 months of gestation.

Keywords

References

  1. Hamilton JA. Colony-stimulating factors in inflammation and autoimmunity. Nat Rev Immunol. 2008;8:533-44. https://doi.org/10.1038/nri2356
  2. Mehta HM, Malandra M, Corey SJ. G-CSF and GM-CSF in neutropenia. J Immunol. 2015;195:1341-9. https://doi.org/10.4049/jimmunol.1500861
  3. Stosser S, Schweizerhof M, Kuner R. Hematopoietic colony-stimulating factors: new players in tumor-nerve interactions. J Mol Med (Berl). 2011;89:321-9. https://doi.org/10.1007/s00109-010-0697-z
  4. Page AV, Liles WC. Colony-stimulating factors in the prevention and management of infectious diseases. Infect Dis Clin North Am. 2011;25:803-17. https://doi.org/10.1016/j.idc.2011.07.007
  5. Melo EO, Canavessi AM, Franco MM, Rumpf R. Animal transgenesis: state of the art and applications. J Appl Genet. 2007;48:47-61. https://doi.org/10.1007/BF03194657
  6. Tanaka H, Tanaka Y, Shinagawa K, Yamagishi Y, Ohtaki K, Asano K. Three types of recombinant human granulocyte colony-stimulating factor have equivalent biological activities in monkeys. Cytokine. 1997;9:360-9. https://doi.org/10.1006/cyto.1996.0177
  7. Bertolini LR, Meade H, Lazzarotto CR, Martins LT, Tavares KC, Bertolini M, et al. The transgenic animal platform for biopharmaceutical production. Transgenic Res. 2016;25:329-43. https://doi.org/10.1007/s11248-016-9933-9
  8. Inoue H, Nojima H, Okayama H. High efficiency transformation of Escherichia coli with plasmids. Gene. 1990;96:23-8. https://doi.org/10.1016/0378-1119(90)90336-P
  9. Sambrook J. Molecular cloning: a laboratory manual. 2001. vol Accessed from. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory; 2001. http://nla.gov.au/nla.cat-vn2284148 Accessed 18 Mar 2019.
  10. Melo EO, Sousa RV, Iguma LT, Franco MM, Rech EL, Rumpf R. Isolation of transfected fibroblast clones for use in nuclear transfer and transgene detection in cattle embryos. Genet Mol Res. 2005;4:812-21.
  11. Cibelli JB, Stice SL, Golueke PJ, Kane JJ, Jerry J, Blackwell C, et al. Cloned transgenic calves produced from nonquiescent fetal fibroblasts. Science. 1998;280:1256-8. https://doi.org/10.1126/science.280.5367.1256
  12. Iguma LT, Lisauskas SF, Melo EO, Franco MM, Pivato I, Vianna GR, et al. Development of bovine embryos reconstructed by nuclear transfer of transfected and non-transfected adult fibroblast cells. Genet Mol Res. 2005;4:55-66.
  13. Holm P, Booth PJ, Schmidt MH, Greve T, Callesen H. High bovine blastocyst development in a static in vitro production system using SOFaa medium supplemented with sodium citrate and myo-inositol with or without serum-proteins. Theriogenology. 1999;52:683-700. https://doi.org/10.1016/S0093-691X(99)00162-4
  14. Kuwayama M, Vajta G, Kato O, Leibo SP. Highly efficient vitrification method for cryopreservation of human oocytes. Reprod Biomed Online. 2005;11:300-8. https://doi.org/10.1016/S1472-6483(10)60837-1
  15. Darzynkiewicz Z. Nucleic acid analysis. Current protocols in cytometry. New York, NY: John Wiley & Sons; 1997. doi:10.1002/0471142956.cy0700s47. Accessed 18 Mar 2019
  16. Verma G, Arora JS, Sethi RS, Mukhopadhyay CS, Verma R. Handmade cloning: recent advances, potential and pitfalls. J Anim Sci Biotechnol. 2015;6:43. https://doi.org/10.1186/s40104-015-0043-y
  17. Blelloch R, Wang Z, Meissner A, Pollard S, Smith A, Jaenisch R. Reprogramming efficiency following somatic cell nuclear transfer is influenced by the differentiation and methylation state of the donor nucleus. Stem Cells. 2006:24;2007-13. https://doi.org/10.1634/stemcells.2006-0050
  18. Tani T, Kato Y, Tsunoda Y. Direct exposure of chromosomes to nonactivated ovum cytoplasm is effective for bovine somatic cell nucleus reprogramming. Biol Reprod. 2001:64;324-30. https://doi.org/10.1095/biolreprod64.1.324
  19. Ogura A, Inoue K, Wakayama T. Recent advancements in cloning by somatic cell nuclear transfer. Philos Trans R Soc Lond B Biol Sci. 2013;368:20110329.
  20. Whitworth KM, Prather RS. Somatic cell nuclear transfer efficiency: how can it be improved through nuclear remodeling and reprogramming? Mol Reprod Dev. 2010;77:1001-15. https://doi.org/10.1002/mrd.21242
  21. Chavatte-Palmer P, Camous S, Jammes H, Le Cleac'h N, Guillomot M, Lee RS. Review: Placental perturbations induce the developmental abnormalities often observed in bovine somatic cell nuclear transfer. Placenta. 2012;33 Suppl:S99-104. https://doi.org/10.1016/j.placenta.2011.09.012
  22. Miglino MA, Pereira FT, Visintin JA, Garcia JM, Meirelles FV, Rumpf R, et al. Placentation in cloned cattle: structure and microvascular architecture. Theriogenology. 2007;68:604-17. https://doi.org/10.1016/j.theriogenology.2007.04.060

Cited by

  1. Strategy to Establish Embryo-Derived Pluripotent Stem Cells in Cattle vol.22, pp.9, 2019, https://doi.org/10.3390/ijms22095011
  2. Effects of Donor Cell Types on the Development of Bovine Embryos Using Cytoplasm Injection Cloning Technology vol.22, pp.11, 2019, https://doi.org/10.3390/ijms22115841