Browse > Article
http://dx.doi.org/10.5187/jast.2019.61.2.61

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)
Publication Information
Journal of Animal Science and Technology / v.61, no.2, 2019 , pp. 61-68 More about this Journal
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
Biotechnology; Genetically modified organisms; Leukopenia; Lipofection;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Hamilton JA. Colony-stimulating factors in inflammation and autoimmunity. Nat Rev Immunol. 2008;8:533-44.   DOI
2 Mehta HM, Malandra M, Corey SJ. G-CSF and GM-CSF in neutropenia. J Immunol. 2015;195:1341-9.   DOI
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.   DOI
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.   DOI
5 Melo EO, Canavessi AM, Franco MM, Rumpf R. Animal transgenesis: state of the art and applications. J Appl Genet. 2007;48:47-61.   DOI
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.   DOI
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.   DOI
8 Inoue H, Nojima H, Okayama H. High efficiency transformation of Escherichia coli with plasmids. Gene. 1990;96:23-8.   DOI
9 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.
10 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.
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.   DOI
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.   DOI
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.   DOI
15 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.   DOI
16 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
17 Verma G, Arora JS, Sethi RS, Mukhopadhyay CS, Verma R. Handmade cloning: recent advances, potential and pitfalls. J Anim Sci Biotechnol. 2015;6:43.   DOI
18 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.   DOI
19 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.   DOI
20 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.
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.   DOI
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.   DOI