Production of transgenic cattle by somatic cell nuclear transfer (SCNT) with the human granulocyte colony-stimulation factor (hG-CSF).
Bruno P CarvalhoAndrielle Thainar Mendes CunhaBianca Damiani Marques SilvaRegivaldo V SousaLigiane de Oliveira LemeMargot Alves Nunes DodeEduardo de Oliveira MeloPublished in: Journal of animal science and technology (2019)
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.
Keyphrases
- fluorescent probe
- induced apoptosis
- cell cycle
- aqueous solution
- bone marrow
- cell cycle arrest
- living cells
- endothelial cells
- extracellular matrix
- copy number
- genome wide
- cell proliferation
- signaling pathway
- poor prognosis
- endoplasmic reticulum stress
- mesenchymal stem cells
- stem cells
- preterm infants
- cell therapy
- induced pluripotent stem cells
- gene expression
- wastewater treatment
- dna methylation
- long non coding rna