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通过体细胞核移植(SCNT)和人粒细胞集落刺激因子(hG-CSF)生产转基因牛。

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

作者信息

Carvalho Bruno P, Cunha Andrielle T M, Silva Bianca D M, Sousa Regivaldo V, Leme Ligiane O, Dode Margot A N, Melo Eduardo O

机构信息

Institute of Biology, Universidade de Brasília, Brasilia-DF, 70910-900, Brazil.

Laboratory of Animal Reproduction, Embrapa Genetic Resources and Biotechnology, Brasília-DF, 70770-917, Brazil.

出版信息

J Anim Sci Technol. 2019 Mar;61(2):61-68. doi: 10.5187/jast.2019.61.2.61. Epub 2019 Mar 31.

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.

摘要

人粒细胞集落刺激因子(hG-CSF)是一种生长和刺激因子,能够诱导骨髓细胞、多种白细胞以及其他造血组织细胞的增殖。hG-CSF用于治疗导致循环白细胞数量减少的异常情况,这可能会损害患者的免疫防御功能。基于这些原因,利用转基因动物的乳腺在生物反应器系统中生产hG-CSF,有可能提升牛遗传物质的价值,并降低制药行业中hG-CSF的生产成本。在本研究中,我们旨在通过用hG-CSF表达盒对牛原代成纤维细胞进行脂质体转染,并通过体细胞核移植(SCNT)技术克隆这些成纤维细胞,从而生产转基因hG-CSF牛。用hG-CSF盒转染的牛成纤维细胞将该构建体稳定插入其基因组,并有效地同步到G0/G1细胞周期阶段。通过SCNT克隆转基因成纤维细胞,产生了103个移植胚胎和2次妊娠,其中一次妊娠达到了7个月。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b46/6582935/4f5f998f7f16/jast-61-2-61-g1.jpg

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